Methods of applying a biological composition to an individual

Abstract

Provided are methods of applying biological compositions, that is, autologous bioadhesive sealant compositions containing one or more biological agents, to an individual, wherein all the blood components used in preparing the composition are derived from the patient who is to receive the biological composition. In one embodiment, the method comprises obtaining a whole blood sample from an individual; forming an inactive platelet rich plasma from the whole blood sample; mixing a biological agent into the inactive platelet rich plasma; obtaining thrombin from the whole blood sample; mixing the thrombin into the inactive platelet rich plasma to form a biological composition; and applying the biological composition to the individual.

The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:  
     
         1 . A method of applying a biological composition to an individual, comprising: 
 obtaining a whole blood sample from an individual;    forming an inactive platelet rich plasma from the whole blood sample;    mixing a biological agent into the inactive platelet rich plasma;    obtaining thrombin from the whole blood sample;    mixing the thrombin into the inactive platelet rich plasma to form a biological composition; and    applying the biological composition to the individual.    
     
     
         2 . The method of  claim 1  further comprising obtaining the biological agent from the individual.  
     
     
         3 . The method of  claim 2  wherein the biological agent is bone.  
     
     
         4 . The method of  claim 1  wherein the biological agent is selected from the group consisting of analgesic compounds, antibacterial compounds, antibiotics, antifungal compounds, anti-inflammatories, antiparasitic compounds, antiviral compounds, enzymes, enzyme inhibitors, glycoproteins, growth factors, hormones, steroids, glucocorticosteroids, immunomodulators, immunoglobulins, minerals, neuroleptics, proteins, peptides, lipoproteins, tumoricidal compounds, tumorstatic compounds, toxins and vitamins.  
     
     
         5 . The method of  claim 1  wherein the biological agent is selected from the group consisting of growth factors, lymphokines and cytokines.  
     
     
         6 . The method of  claim 1  wherein the biological agent is selected from the group consisting of antibacterial compounds, bacteriocidal compounds and bacteriostatic compounds.  
     
     
         7 . The method of  claim 1  wherein the biological agent is selected from the group consisting of antibiotics, adriamycin, erythromycin, gentimycin, penicillin and tobramycin.  
     
     
         8 . The method of  claim 1  wherein the biological agent is selected from the group consisting of vitamins, Vitamin A, Vitamin E, Vitamin B, Vitamin C and Vitamin D.  
     
     
         9 . The method of  claim 1  wherein the biological agent is cells.  
     
     
         10 . The method of  claim 9  wherein the cells are obtained from the individual.  
     
     
         11 . The method of  claim 10  wherein the cells are bone cells.  
     
     
         12 . The method of  claim 1  wherein the step of applying the biological composition to the individual includes applying the biological composition to a wound.  
     
     
         13 . The method of  claim 1  further comprising: 
 forming a first and second portion of the inactive platelet rich plasma;  
 reactivating the first portion of the inactive platelet rich plasma to form a clot;  
 triturating the clot to obtain a serum comprising the thrombin;  
 mixing the biological agent into the second portion of the inactive platelet rich plasma; and  
 mixing the serum into the inactive platelet rich plasma to form the biological composition.  
 
     
     
         14 . The method of  claim 1  further comprising: 
 forming a first and second portion of the inactive platelet rich plasma;  
 reactivating the first portion of the inactive platelet rich plasma to form a clot;  
 squeezing the clot through a mesh to obtain a serum comprising the thrombin;  
 mixing the biological agent into the second portion of the inactive platelet rich plasma; and  
 mixing the serum into the inactive platelet rich plasma to form the biological composition.  
 
     
     
         15 . The method of  claim 1  further comprising: 
 forming a first and second portion of the inactive platelet rich plasma;  
 reactivating the first portion of the inactive platelet rich plasma to form a clot;  
 centrifuging the clot to obtain a serum comprising the thrombin;  
 mixing the biological agent into the second portion of the inactive platelet rich plasma; and  
 mixing the serum into the inactive platelet rich plasma to form the biological composition.  
 
     
     
         16 . The method of  claim 1  wherein the step of applying the biological composition to the individual results in a barrier against loss of blood from a vessel.  
     
     
         17 . The method of  claim 1  wherein the step of applying the biological composition to the individual results in a scaffold for repair of tissue.  
     
     
         18 . The method of  claim 1  wherein the step of applying the biological composition to the individual includes applying the biological composition to a needle hole.  
     
     
         19 . The method of  claim 1  wherein the step of applying the biological composition to the individual includes applying the biological composition to an arterial tear.  
     
     
         20 . The method of  claim 1  wherein the step of applying the biological composition to the individual includes applying the biological composition to tissue.  
     
     
         21 . The method of  claim 1  wherein the step of applying the biological composition to the individual includes applying the biological composition to bone.  
     
     
         22 . The method of  claim 1  wherein the step of applying the biological composition to the individual includes applying the biological composition to a nerve.  
     
     
         23 . The method of  claim 1  wherein the step of applying the biological composition to the individual includes applying the biological composition to a vessel.  
     
     
         24 . The method of  claim 1  wherein the whole blood sample obtained from the individual is anticoagulated.  
     
     
         25 . The method of  claim 24  wherein the whole blood sample is mixed with an anticoagulation agent.  
     
     
         26 . The method of  claim 25  wherein the anticoagulation agent is sodium citrate.  
     
     
         27 . The method of  claim 25  wherein the anticoagulation agent is a chelating agent.  
     
     
         28 . The method of  claim 25  wherein the anticoagulation agent is heparin.  
     
     
         29 . The method of  claim 1  wherein the inactive platelet rich plasma is formed by centrifuging the whole blood sample.  
     
     
         30 . The method of  claim 1  further comprising: 
 dividing the inactive platelet rich plasma into a first portion and a second portion;  
 mixing the first portion with a restoration agent to form a clot;  
 triturating the clot to obtain the thrombin; and  
 mixing the thrombin into the second portion of the inactive platelet rich plasma to form the biological composition.  
 
     
     
         31 . The method of  claim 30  wherein the restoration agent is calcium chloride.  
     
     
         32 . The method of  claim 30  wherein the restoration agent is calcium gluconate.  
     
     
         33 . The method of  claim 30  wherein the restoration agent is heparinase.  
     
     
         34 . The method of  claim 1  wherein the biological composition is applied to a cardiovascular locale.  
     
     
         35 . The method of  claim 1  wherein the biological composition is applied to a thoracic locale.  
     
     
         36 . The method of  claim 1  wherein the biological composition is applied to a transplantation locale.  
     
     
         37 . The method of  claim 1  wherein the biological composition is applied to a head locale.  
     
     
         38 . The method of  claim 1  wherein the biological composition is applied to a neck locale.  
     
     
         39 . The method of  claim 1  wherein the biological composition is applied to an oral locale.  
     
     
         40 . The method of  claim 1  wherein the biological composition is applied to a gastrointestinal locale.  
     
     
         41 . The method of  claim 1  wherein the biological composition is applied to an orthopedic locale.  
     
     
         42 . The method of  claim 1  wherein the biological composition is applied to a neurosurgical locale.  
     
     
         43 . The method of  claim 1  wherein the biological composition is applied to a plastic surgery locale.  
     
     
         44 . The method of  claim 1  further comprising: 
 forming an inactive platelet poor plasma from the whole blood sample;  
 reactivating the inactive platelet poor plasma to form a clot;  
 triturating the clot to obtain a serum comprising the thrombin; and  
 mixing the serum into the inactive platelet rich plasma to form the biological composition.  
 
     
     
         45 . The method of  claim 44  wherein triturating the clot includes squeezing the clot through a mesh to obtain the serum comprising the thrombin.  
     
     
         46 . The method of  claim 44  wherein triturating the clot includes centrifuging the clot to obtain the serum comprising the thrombin.  
     
     
         47 . The method of  claim 44  wherein the inactive platelet poor plasma is formed by centrifuging the whole blood sample.  
     
     
         48 . The method of  claim 44  wherein reactivating the inactive platelet poor plasma includes mixing the inactive platelet poor plasma with a restoration agent.  
     
     
         49 . The method of  claim 48  wherein the restoration agent is calcium chloride.  
     
     
         50 . The method of  claim 48  wherein the restoration agent is calcium gluconate.  
     
     
         51 . The method of  claim 48  wherein the restoration agent is heparinase.  
     
     
         52 . A method of applying a biological composition to an individual, comprising: 
 obtaining a whole blood sample from an individual;    forming an inactive platelet poor plasma from the whole blood sample;    mixing a biological agent into the inactive platelet poor plasma;    obtaining thrombin from the whole blood sample;    mixing the thrombin into the inactive platelet poor plasma to form a biological composition; and    applying the biological composition to the individual.    
     
     
         53 . The method of  claim 52  further comprising obtaining the biological agent from the individual.  
     
     
         54 . The method of  claim 53  wherein the biological agent is bone.  
     
     
         55 . The method of  claim 52  wherein the biological agent is selected from the group consisting of analgesic compounds, antibacterial compounds, antibiotics, antifungal compounds, anti-inflammatories, antiparasitic compounds, antiviral compounds, enzymes, enzyme inhibitors, glycoproteins, growth factors, hormones, steroids, glucocorticosteroids, immunomodulators, immunoglobulins, minerals, neuroleptics, proteins, peptides, lipoproteins, tumoricidal compounds, tumorstatic compounds, toxins and vitamins.  
     
     
         56 . The method of  claim 52  wherein the biological agent is selected from the group consisting of growth factors, lymphokines and cytokines.  
     
     
         57 . The method of  claim 52  wherein the biological agent is selected from the group consisting of antibacterial compounds, bacteriocidal compounds and bacteriostatic compounds.  
     
     
         58 . The method of  claim 52  wherein the biological agent is selected from the group consisting of antibiotics, adriamycin, erythromycin, gentimycin, penicillin and tobramycin.  
     
     
         59 . The method of  claim 52  wherein the biological agent is selected from the group consisting of vitamins, Vitamin A, Vitamin E, Vitamin B, Vitamin C and Vitamin D.  
     
     
         60 . The method of  claim 52  wherein the biological agent is cells.  
     
     
         61 . The method of  claim 60  wherein the cells are obtained from the individual.  
     
     
         62 . The method of  claim 61  wherein the cells are bone cells.  
     
     
         63 . The method of  claim 52  wherein the step of applying the biological composition to the individual includes applying the biological composition to a wound.  
     
     
         64 . The method of  claim 52  further comprising: 
 forming a first and second portion of the inactive platelet poor plasma;  
 reactivating the first portion of the inactive platelet poor plasma to form a clot;  
 triturating the clot to obtain a serum comprising the thrombin;  
 mixing the biological agent into the second portion of the inactive platelet poor plasma; and  
 mixing the serum into the inactive platelet poor plasma to form the biological composition.  
 
     
     
         65 . The method of  claim 52  further comprising: 
 forming a first and second portion of the inactive platelet poor plasma;  
 reactivating the first portion of the inactive platelet poor plasma to form a clot;  
 squeezing the clot through a mesh to obtain a serum comprising the thrombin;  
 mixing the biological agent into the second portion of the inactive platelet poor plasma; and  
 mixing the serum into the inactive platelet poor plasma to form the biological composition.  
 
     
     
         66 . The method of  claim 52  further comprising: 
 forming a first and second portion of the inactive platelet poor plasma;  
 reactivating the first portion of the inactive platelet poor plasma to form a clot;  
 centrifuging the clot to obtain a serum comprising the thrombin;  
 mixing the biological agent into the second portion of the inactive platelet poor plasma; and  
 mixing the serum into the inactive platelet poor plasma to form the biological composition.  
 
     
     
         67 . The method of  claim 52  wherein the step of applying the biological composition to the individual results in a barrier against loss of blood from a vessel.  
     
     
         68 . The method of  claim 52  wherein the step of applying the biological composition to the individual results in a scaffold for repair of tissue.  
     
     
         69 . The method of  claim 52  wherein the step of applying the biological composition to the individual includes applying the biological composition to a needle hole.  
     
     
         70 . The method of  claim 52  wherein the step of applying the biological composition to the individual includes applying the biological composition to an arterial tear.  
     
     
         71 . The method of  claim 52  wherein the step of applying the biological composition to the individual includes applying the biological composition to tissue.  
     
     
         72 . The method of  claim 52  wherein the step of applying the biological composition to the individual includes applying the biological composition to bone.  
     
     
         73 . The method of  claim 52  wherein the step of applying the biological composition to the individual includes applying the biological composition to a nerve.  
     
     
         74 . The method of  claim 52  wherein the step of applying the biological composition to the individual includes applying the biological composition to a vessel.  
     
     
         75 . The method of  claim 52  wherein the whole blood sample obtained from the individual is anticoagulated.  
     
     
         76 . The method of  claim 75  wherein the whole blood sample is mixed with an anticoagulation agent.  
     
     
         77 . The method of  claim 76  wherein the anticoagulation agent is sodium citrate.  
     
     
         78 . The method of  claim 76  wherein the anticoagulation agent is a chelating agent.  
     
     
         79 . The method of  claim 76  wherein the anticoagulation agent is heparin.  
     
     
         80 . The method of  claim 52  wherein the inactive platelet poor plasma is formed by centrifuging the whole blood sample.  
     
     
         81 . The method of  claim 52  further comprising: 
 dividing the inactive platelet poor plasma into a first portion and a second portion;  
 mixing the first portion with a restoration agent to form a clot;  
 triturating the clot to obtain the thrombin; and  
 mixing the thrombin into the second portion of the inactive platelet poor plasma to form the biological composition.  
 
     
     
         82 . The method of  claim 81  wherein the restoration agent is calcium chloride.  
     
     
         83 . The method of  claim 81  wherein the restoration agent is calcium gluconate.  
     
     
         84 . The method of  claim 81  wherein the restoration agent is heparinase.  
     
     
         85 . The method of  claim 52  wherein the biological composition is applied to a cardiovascular locale.  
     
     
         86 . The method of  claim 52  wherein the biological composition is applied to a thoracic locale.  
     
     
         87 . The method of  claim 52  wherein the biological composition is applied to a transplantation locale.  
     
     
         88 . The method of  claim 52  wherein the biological composition is applied to a head locale.  
     
     
         89 . The method of  claim 52  wherein the biological composition is applied to a neck locale.  
     
     
         90 . The method of  claim 52  wherein the biological composition is applied to an oral locale.  
     
     
         91 . The method of  claim 52  wherein the biological composition is applied to a gastrointestinal locale.  
     
     
         92 . The method of  claim 52  wherein the biological composition is applied to an orthopedic locale.  
     
     
         93 . The method of  claim 52  wherein the biological composition is applied to a neurosurgical locale.  
     
     
         94 . The method of  claim 52  wherein the biological composition is applied to a plastic surgery locale.  
     
     
         95 . The method of  claim 52  further comprising: 
 forming an inactive platelet rich plasma from the whole blood sample;  
 reactivating the inactive platelet rich plasma to form a clot;  
 triturating the clot to obtain a serum comprising the thrombin; and  
 mixing the serum into the inactive platelet poor plasma to form the biological composition.  
 
     
     
         96 . The method of  claim 95  wherein triturating the clot includes squeezing the clot through a mesh to obtain the serum comprising the thrombin.  
     
     
         97 . The method of  claim 95  wherein triturating the clot includes centrifuging the clot to obtain the serum comprising the thrombin.  
     
     
         98 . The method of  claim 95  wherein the inactive platelet rich plasma is formed by centrifuging the whole blood sample.  
     
     
         99 . The method of  claim 95  wherein reactivating the inactive platelet rich plasma includes mixing the inactive platelet rich plasma with a restoration agent.  
     
     
         100 . The method of  claim 99  wherein the restoration agent is calcium chloride.  
     
     
         101 . The method of  claim 99  wherein the restoration agent is calcium gluconate.  
     
     
         102 . The method of  claim 99  wherein the restoration agent is heparinase.  
     
     
         103 . A method of applying a biological composition to a patient, comprising: 
 obtaining a whole blood sample from the patient;    mixing an anticoagulation agent into the whole blood sample;    centrifuging the anticoagulated whole blood sample to form an inactive platelet rich plasma;    dividing the inactive platelet rich plasma into a first portion and a second portion;    mixing a restoration agent into the first portion of the inactive platelet rich plasma to form a clot;    triturating the clot to obtain a serum comprising thrombin;    mixing a biological agent into the second portion of the inactive platelet rich plasma; and    mixing the serum into the inactive platelet rich plasma to form the biological composition; and    applying the biological composition to a desired patient locale where it congeals.    
     
     
         104 . The method of  claim 103  further comprising obtaining the biological agent from the patient.  
     
     
         105 . The method of  claim 104  wherein the biological agent is bone.  
     
     
         106 . The method of  claim 103  wherein the biological agent is selected from the group consisting of analgesic compounds, antibacterial compounds, antibiotics, antifungal compounds, anti-inflammatories, antiparasitic compounds, antiviral compounds, enzymes, enzyme inhibitors, glycoproteins, growth factors, hormones, steroids, glucocorticosteroids, immunomodulators, immunoglobulins, minerals, neuroleptics, proteins, peptides, lipoproteins, tumoricidal compounds, tumorstatic compounds, toxins and vitamins.  
     
     
         107 . The method of  claim 103  wherein the biological agent is selected from the group consisting of growth factors, lymphokines and cytokines.  
     
     
         108 . The method of  claim 103  wherein the biological agent is selected from the group consisting of antibacterial compounds, bacteriocidal compounds and bacteriostatic compounds.  
     
     
         109 . The method of  claim 103  wherein the biological agent is selected from the group consisting of antibiotics, adriamycin, erythromycin, gentimycin, penicillin and tobramycin.  
     
     
         110 . The method of  claim 103  wherein the biological agent is selected from the group consisting of vitamins, Vitamin A, Vitamin E, Vitamin B, Vitamin C and Vitamin D.  
     
     
         111 . The method of  claim 103  wherein the biological agent is cells.  
     
     
         112 . The method of  claim 111  wherein the cells are obtained from the patient.  
     
     
         113 . The method of  claim 112  wherein the cells are bone cells.  
     
     
         114 . The method of  claim 103  wherein the step of applying the biological composition includes applying the biological composition to a wound.  
     
     
         115 . The method of  claim 103  wherein the step of applying the biological composition results in a barrier against loss of blood from a vessel.  
     
     
         116 . The method of  claim 103  wherein the step of applying the biological composition results in a scaffold for repair of tissue.  
     
     
         117 . The method of  claim 103  wherein the step of applying the biological composition includes applying the biological composition to a needle hole.  
     
     
         118 . The method of  claim 103  wherein the step of applying the biological composition includes applying the biological composition to an arterial tear.  
     
     
         119 . The method of  claim 103  wherein the step of applying the biological composition includes applying the biological composition to tissue.  
     
     
         120 . The method of  claim 103  wherein the step of applying the biological composition includes applying the biological composition to bone.  
     
     
         121 . The method of  claim 103  wherein the step of applying the biological composition includes applying the biological composition to a nerve.  
     
     
         122 . The method of  claim 103  wherein the step of applying the biological composition includes applying the biological composition to a vessel.  
     
     
         123 . The method of  claim 103  wherein the anticoagulation agent is sodium citrate.  
     
     
         124 . The method of  claim 103  wherein the anticoagulation agent is a chelating agent.  
     
     
         125 . The method of  claim 103  wherein the anticoagulation agent is heparin.  
     
     
         126 . The method of  claim 103  wherein the restoration agent is calcium chloride.  
     
     
         127 . The method of  claim 103  wherein the restoration agent is calcium gluconate.  
     
     
         128 . The method of  claim 103  wherein the restoration agent is heparinase.  
     
     
         129 . The method of  claim 103  wherein the biological composition is applied to a cardiovascular locale.  
     
     
         130 . The method of  claim 103  wherein the biological composition is applied to a thoracic locale.  
     
     
         131 . The method of  claim 103  wherein the biological composition is applied to a transplantation locale.  
     
     
         132 . The method of  claim 103  wherein the biological composition is applied to a head locale.  
     
     
         133 . The method of  claim 103  wherein the biological composition is applied to a neck locale.  
     
     
         134 . The method of  claim 103  wherein the biological composition is applied to an oral locale.  
     
     
         135 . The method of  claim 103  wherein the biological composition is applied to a gastrointestinal locale.  
     
     
         136 . The method of  claim 103  wherein the biological composition is applied to an orthopedic locale.  
     
     
         137 . The method of  claim 103  wherein the biological composition is applied to a neurosurgical locale.  
     
     
         138 . The method of  claim 103  wherein the biological composition is applied to a plastic surgery locale.  
     
     
         139 . The method of  claim 103  wherein triturating the clot includes squeezing the clot through a mesh to obtain the serum comprising the thrombin.  
     
     
         140 . The method of  claim 103  wherein triturating the clot includes centrifuging the clot to obtain the serum comprising the thrombin.  
     
     
         141 . A method of applying a biological composition to a patient, comprising: 
 obtaining an inactive platelet rich plasma;    mixing a biological agent into the inactive platelet rich plasma;    mixing human recombinant thromboplastin into the inactive platelet rich plasma to form the biological composition; and    applying the biological composition to a desired patient locale.    
     
     
         142 . The method of  claim 141  further comprising obtaining the inactive platelet rich plasma from the patient.  
     
     
         143 . The method of  claim 141  further comprising obtaining the biological agent from the patient.  
     
     
         144 . The method of  claim 141  wherein the biological agent is bone.  
     
     
         145 . The method of  claim 141  wherein the biological agent is selected from the group consisting of analgesic compounds, antibacterial compounds, antibiotics, antifungal compounds, anti-inflammatories, antiparasitic compounds, antiviral compounds, enzymes, enzyme inhibitors, glycoproteins, growth factors, hormones, steroids, glucocorticosteroids, immunomodulators, immunoglobulins, minerals, neuroleptics, proteins, peptides, lipoproteins, tumoricidal compounds, tumorstatic compounds, toxins and vitamins.  
     
     
         146 . The method of  claim 141  wherein the biological agent is selected from the group consisting of growth factors, lymphokines and cytokines.  
     
     
         147 . The method of  claim 141  wherein the biological agent is selected from the group consisting of antibacterial compounds, bacteriocidal compounds and bacteriostatic compounds.  
     
     
         148 . The method of  claim 141  wherein the biological agent is selected from the group consisting of antibiotics, adriamycin, erythromycin, gentimycin, penicillin and tobramycin.  
     
     
         149 . The method of  claim 141  wherein the biological agent is selected from the group consisting of vitamins, Vitamin A, Vitamin E, Vitamin B, Vitamin C and Vitamin D.  
     
     
         150 . The method of  claim 141  wherein the biological agent is cells.  
     
     
         151 . The method of  claim 150  wherein the cells are obtained from the patient.  
     
     
         152 . The method of  claim 151  wherein the cells are bone cells.  
     
     
         153 . The method of  claim 141  wherein the step of applying the biological composition includes applying the biological composition to a wound.  
     
     
         154 . The method of  claim 141  wherein the step of applying the biological composition results in a barrier against loss of blood from a vessel.  
     
     
         155 . The method of  claim 141  wherein the step of applying the biological composition results in a scaffold for repair of tissue.  
     
     
         156 . The method of  claim 141  wherein the step of applying the biological composition includes applying the biological composition to a needle hole.  
     
     
         157 . The method of  claim 141  wherein the step of applying the biological composition includes applying the biological composition to an arterial tear.  
     
     
         158 . The method of  claim 141  wherein the step of applying the biological composition includes applying the biological composition to tissue.  
     
     
         159 . The method of  claim 141  wherein the step of applying the biological composition includes applying the biological composition to bone.  
     
     
         160 . The method of  claim 141  wherein the step of applying the biological composition includes applying the biological composition to a nerve.  
     
     
         161 . The method of  claim 141  wherein the step of applying the biological composition includes applying the biological composition to a vessel.  
     
     
         162 . The method of  claim 141  wherein the desired patient locale is a cardiovascular locale.  
     
     
         163 . The method of  claim 141  wherein the desired patient locale is a thoracic locale.  
     
     
         164 . The method of  claim 141  wherein the desired patient locale is a transplantation locale.  
     
     
         165 . The method of  claim 141  wherein the desired patient locale is a head locale.  
     
     
         166 . The method of  claim 141  wherein the desired patient locale is a neck locale.  
     
     
         167 . The method of  claim 141  wherein the desired patient locale is an oral locale.  
     
     
         168 . The method of  claim 141  wherein the desired patient locale is a gastrointestinal locale.  
     
     
         169 . The method of  claim 141  wherein the desired patient locale is an orthopedic locale.  
     
     
         170 . The method of  claim 141  wherein the desired patient locale is a neurosurgical locale.  
     
     
         171 . The method of  claim 141  wherein the desired patient locale is a plastic surgery locale.  
     
     
         172 . A method of applying a biological composition to an individual, comprising: 
 obtaining an anticoagulated whole blood sample from an individual;    forming an inactive platelet rich plasma from the whole blood sample;    dividing the inactive platelet rich plasma into first and second portions;    mixing a biological agent into the first portion of the inactive platelet rich plasma;    obtaining autologous thrombin from the second portion of the inactive platelet rich plasma;    mixing the autologous thrombin with the inactive platelet rich plasma containing the biological agent to form a biological composition; and    applying the biological composition to the individual.    
     
     
         173 . A method of applying a biological composition to an individual, comprising: 
 obtaining an anticoagulated whole blood sample from an individual;    forming an inactive platelet poor plasma from the whole blood sample;    dividing the inactive platelet poor plasma into first and second portions;    mixing a biological agent with the first portion of the inactive platelet poor plasma;    obtaining autologous thrombin from the second portion of the inactive platelet poor plasma;    mixing the autologous thrombin with the inactive platelet poor plasma containing the biological agent to form a biological composition; and    applying the biological composition to the individual.