探討豬源血小板裂解液 於紐西蘭白兔後腿膝關節腔之生物相容性

Exploring the biocompatibility of porcine platelet lysate in the hind leg knee joint cavity of New Zealand White rabbits

朱依倢(YI-CHIEH CHU)

指導教授 ： 謝明發

中原大學/工學院/生物醫學工程學系/碩士(2023年)

https://doi.org/10.6840/cycu202301738

摘要

以離心技術分離血球且純化獲得高濃度血小板血漿，稱之為血小板濃厚血漿，其經凍融循環獲得一種淺黃色澄清液體稱之為血小板裂解液，其中含有豐富的生長因子、蛋白質與少量抗體。血小板裂解液具有促進癒合的作用，目前多使用於再生醫學，如口腔外科、關節軟骨再生和軟組織癒合等。本研究探討豬隻血小板裂解液的製程方法及其注射於兔子後腿關節腔後的生物相容性。實驗使用兩種不同豬隻來源的血小板濃厚血漿，分別以低張溶液裂解法、超音波震盪裂解法、凍融裂解法評估其裂解條件製備的生長因子劑型。實驗結果顯示，經凍融法四次循環後血小板明顯降低至4 x103/µL；另以酵素結合免疫吸附分析法檢測乙型轉化生長因子濃度，結果顯示凍融裂解法為最為有效之裂解血小板製程方法，可以獲得4.29 ng/mL生長因子。生物相容性試驗是用雄性紐西蘭大白兔膝關節注射0.3 mL血小板裂解液，並觀察組織切片及臨床病理情形，以此判斷注射後的組織反應。結果顯示，注射部位的膝蓋並無紅腫熱之發炎情形，且動物的活動保持正常、體重均增加。故異種血小板裂解液具有應用於不同物種之傷口修復之潛力。

關鍵字

血小板裂解液 ； 豬源血小板濃厚液 ； 再生醫學 ； 生物相容性

並列摘要

The plasma with a high concentration of platelets obtained by centrifugation and purification is called platelet-rich plasma. The light-yellow transparent liquid obtained by the freeze-thaw cycle is called platelet lysate (PL), which is rich in growth factors, proteins, and a small number of antibodies. PL has healing-promoting effects and is currently used in regenerative medicine, such as oral surgery, joint cartilage regeneration, and soft tissue healing. In this study, PL was produced from porcine whole blood, and its production method and biocompatibility after injection into the joint cavity of rabbits were investigated. Two different types of platelet-rich plasma from different pig sources were used to evaluate the platelet lysis conditions and the concentrations of transforming growth factor-beta (TGF-) prepared by hypotonic lysis, ultrasonic shock lysis, and freeze-thaw lysis, respectively. The optimal result among four lysis methods showed that the platelet count decreased to 4 x103/µL after four cycles of freeze-thaw lysis, and the concentration of TGF- produced by different lysis methods was measured by enzyme-linked immunosorbent assay. In the in-vivo biocompatibility test, 0.3 mL of PL was injected into the joint cavity of the knee of male New Zealand rabbits, and the immune response after injection was determined by observing histological sections and clinical pathology. The results showed that there was no redness, swelling, or inflammation of the knee joints, and the rabbits maintained normal activity and increased body weight. Therefore, the allogeneic platelet lysate has the potential to be used for wound repair in different species.

並列關鍵字

platelet lysate ； porcine-derived platelet concentrates ； regenerative medicine ； biocompatibility

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