【摘要】：關(guān)節(jié)軟骨損傷是導(dǎo)致人類運(yùn)動(dòng)功能障礙的重要原因,異體關(guān)節(jié)軟骨移植是一種有效的治療手段。移植手術(shù)的廣泛開展有賴于關(guān)節(jié)軟骨庫存的建立,以及成熟的關(guān)節(jié)軟骨長(zhǎng)期保存技術(shù)。近年來提出的液相線跟蹤法(liquidus-tracking method,LTM)在玻璃化保存關(guān)節(jié)軟骨方面取得了較好的效果,是一項(xiàng)很有應(yīng)用前景的保存方法,對(duì)該方法內(nèi)在機(jī)理的深入研究將有助于保存方案的完善以及保存效果的提高。本文全面綜述了國(guó)內(nèi)外關(guān)節(jié)軟骨長(zhǎng)期保存的歷史和研究現(xiàn)狀,以羊膝關(guān)節(jié)軟骨為對(duì)象,以減少關(guān)節(jié)軟骨在低溫保護(hù)劑中的處理步數(shù)和處理時(shí)間為目標(biāo),從傳熱傳質(zhì)的角度對(duì)LTM中低溫保護(hù)劑處理過程進(jìn)行了理論和實(shí)驗(yàn)研究,開展了以下工作： 1)實(shí)驗(yàn)確定了關(guān)節(jié)軟骨內(nèi)需要載入的二甲亞砜最佳濃度。先采用差示掃描量熱技術(shù)獲得了不同濃度(43%(w/w)～50%(w/w))的二甲亞砜/CPTes2溶液的玻璃化臨界冷卻速率和避免反玻璃化臨界升溫速率,隨后對(duì)4mL二甲亞砜/CPTes2溶液的冷卻和升溫速率進(jìn)行了實(shí)驗(yàn)測(cè)量,根據(jù)上述兩項(xiàng)實(shí)驗(yàn)確定可以采用47.5%(w/w)作為關(guān)節(jié)軟骨樣品內(nèi)二甲亞砜的最終載入濃度。 2)實(shí)驗(yàn)研究了二甲亞砜對(duì)關(guān)節(jié)軟骨的滲透性。提出采用紫外-可見吸收光譜法定量分析二甲亞砜的含量并進(jìn)行了實(shí)驗(yàn)驗(yàn)證,隨后應(yīng)用此方法對(duì)寬溫度范圍(37℃～-30℃)內(nèi)不同濃度(10%(w/w)～64.5%(w/w)二甲亞砜滲透關(guān)節(jié)軟骨的過程進(jìn)行了定量研究,獲得了經(jīng)歷不同處理過程的關(guān)節(jié)軟骨片內(nèi)的二甲亞砜的含量。實(shí)驗(yàn)發(fā)現(xiàn),滲透達(dá)平衡時(shí)軟骨片內(nèi)的二甲亞砜濃度約為外部處理溶液濃度的90%。 3)建立了新的描述二甲亞砜滲透關(guān)節(jié)軟骨過程的理論模型。滲透模型可實(shí)現(xiàn)對(duì)二甲亞砜在關(guān)節(jié)軟骨中的有效擴(kuò)散系數(shù)進(jìn)行直接計(jì)算,彌補(bǔ)了現(xiàn)有模型對(duì)實(shí)驗(yàn)數(shù)據(jù)依賴性大的不足。在本文的研究溫度和濃度范圍內(nèi),二甲亞砜在關(guān)節(jié)軟骨內(nèi)的滲透量的模型預(yù)測(cè)結(jié)果與實(shí)驗(yàn)結(jié)果吻合良好。 4)對(duì)關(guān)節(jié)軟骨片的LTM保存方案進(jìn)行了分析和優(yōu)化。運(yùn)用上述新的理論模型,結(jié)合軟骨細(xì)胞膜的傳質(zhì)模型以及軟骨細(xì)胞的生物物理特性,對(duì)現(xiàn)有LTM方案中的傳質(zhì)現(xiàn)象以及軟骨細(xì)胞可能遭受的損傷進(jìn)行了理論分析,并在此基礎(chǔ)上提出了對(duì)階梯式液相線跟蹤法保存關(guān)節(jié)軟骨片的方案進(jìn)行改進(jìn)的思路。按新思路提出的方案在總處理步數(shù)上可減少2步,總處理時(shí)間上可縮短35%。處理步數(shù)和處理時(shí)間的減少不僅可節(jié)省人力物力,而且在處理溫度和處理濃度保持不變的情況下,處理時(shí)間的縮短將有助于降低軟骨細(xì)胞遭受毒性損傷和寒冷損傷的風(fēng)險(xiǎn)。 5)提出了骨軟骨栓的LTM保存方案。對(duì)于階梯式和連續(xù)式LTM,骨軟骨栓的預(yù)計(jì)處理時(shí)間分別為相同厚度軟骨片的3.7倍和3.3倍。
[Abstract]:Articular cartilage injury is an important cause of human motor dysfunction. Allogenic articular cartilage transplantation is an effective treatment. The extensive development of transplantation depends on the establishment of articular cartilage inventory and the long-term preservation of mature articular cartilage. The liquid line tracking method (liquidus-tracking method,LTM) proposed in recent years has achieved good results in vitrification preservation of articular cartilage, and it is a promising preservation method. The further study of the internal mechanism of this method will be helpful to the improvement of the preservation scheme and the improvement of the preservation effect. In this paper, the history and research status of long-term preservation of articular cartilage at home and abroad are reviewed. The sheep knee cartilage is taken as the object, and the goal is to reduce the number of treatment steps and treatment time of articular cartilage in cryopreservation agent. From the point of view of heat and mass transfer, the treatment process of cryogenic protectant in LTM was studied theoretically and experimentally. the following work was carried out: 1) the optimum concentration of dimethyl sulfoxide needed to be loaded into articular cartilage was determined. Firstly, the critical cooling rate of vitrification and the critical heating rate of avoiding antivitrification of dimethyl sulfoxide / CPTes2 solution with different concentrations (43% (w 鈮，

本文編號(hào)：2474386