兩種富血小板纖維蛋白的降解特性研究
發(fā)布時間：2018-08-25 09:16
【摘要】：目的:從抽取人血液制備兩種富血小板纖維蛋白進(jìn)行的體外降解實(shí)驗(yàn)和實(shí)驗(yàn)動物口內(nèi)植入實(shí)驗(yàn)兩個部分對比分析富血小板纖維蛋白和改良型富血小板纖維蛋白(Platelet-Rich Fibrin,PRF和Advanced Platelet-Rich Fibrin,A-PRF)的降解特性,評估其臨床應(yīng)用價值,為A-PRF膜和PRF膜的臨床應(yīng)用提供相關(guān)實(shí)驗(yàn)依據(jù)。方法:1.體外降解實(shí)驗(yàn):10名志愿者各抽取10ml肘靜脈血用以制取A-PRF和PRF膜,A-PRF以Ghanaati法制備(1500rpm,14min),PRF以Choukroun法制備(2700rpm,12min)。制備好的A-PRF、PRF膜浸泡在人工唾液內(nèi)置于二氧化碳培養(yǎng)箱,每天記錄膜的物理狀態(tài)并測量膜標(biāo)本的重量。2.兔口內(nèi)植入實(shí)驗(yàn):36只實(shí)驗(yàn)兔隨機(jī)分為A-PRF組(A組)和PRF組(B組),每只兔抽取10ml血液用以制備A-PRF或PRF膜,A-PRF(2000rpm,24min),PRF(3250rpm,10min)。將制備好的膜修剪成6mm直徑的圓形膜片。于每只兔硬腭處制備兩處模型:在距上頜內(nèi)側(cè)門牙4.5mm的腭中線處用環(huán)切鉆制備直徑5mm的軟組織缺損,剝離粘骨膜顯露骨面,并向周圍潛行分離1mm;在軟組織缺損下方2mm約一個腭皺襞處用手術(shù)刀片做邊長5mm的角形切口,翻全層粘骨膜瓣顯露骨面。A組在軟組織缺損區(qū)植入自體A-PRF圓形膜片,邊緣縫合固定(A1組,即A-PRF膜暴露組);角形瓣內(nèi)植入A-PRF圓形膜片并縫合(A2組,即A-PRF膜埋置組)。B組在兩處植入自體PRF圓形膜片,軟組織缺損區(qū)為B1組(即PRF膜暴露組),角形瓣區(qū)為B2組(即PRF膜埋置組)。四組每組18個實(shí)驗(yàn)樣本。在術(shù)后3d、5d、7d、10d、14d、21d,A、B兩組各隨機(jī)處死3只動物,取出位于上顎部的膜標(biāo)本,進(jìn)行形態(tài)學(xué)觀察并稱重計算降解率,掃描電鏡觀察降解過程,HE染色,觀察組織病理變化,進(jìn)行炎性評分。結(jié)果:1.體外降解實(shí)驗(yàn):(1)在人工唾液培養(yǎng)環(huán)境下,第14天所有PRF膜標(biāo)本均完全降解,第17天所有A-PRF膜標(biāo)本均完全降解。(2)A-PRF和PRF膜的降解速度有差異(P0.05)。2.兔口內(nèi)植入實(shí)驗(yàn):(1)形態(tài)學(xué)觀察:術(shù)后第7d:A2和B2組創(chuàng)口基本愈合;術(shù)后第14d,A1和B1組創(chuàng)口基本愈合。術(shù)后第7d:B1組膜材料明顯吸收較難完整取出;術(shù)后第10d:B1組無可見標(biāo)本,A1組、B2組膜取出困難;術(shù)后第14d:A2組膜取出困難,A1組、B2組無可見標(biāo)本;術(shù)后第21d:A2組無法找到植入標(biāo)本。(2)降解情況:完全降解的時間A1組14天,A2組21天,B1組10天,B2組14天。組間對比:A1、A2兩組膜標(biāo)本的降解率在前5個時間節(jié)點(diǎn)有統(tǒng)計學(xué)差異(P0.05);B1、B2組的降解率除第三天無統(tǒng)計學(xué)差異(P=0.0660.05),其余時間點(diǎn)差異均有統(tǒng)計學(xué)意義(P0.05);A1、B1兩組膜第5天、7天、10天差異有統(tǒng)計學(xué)意義(P0.05);A2、B2組在五個時間節(jié)點(diǎn)的降解率均有統(tǒng)計學(xué)差異(P0.05)。(3)掃描電鏡結(jié)果:A-PRF和PRF膜均呈三維網(wǎng)狀結(jié)構(gòu),由三叉結(jié)構(gòu)連接。降解過程中纖維蛋白網(wǎng)狀結(jié)構(gòu)逐漸崩解,表現(xiàn)為纖維蛋白條索的斷裂,網(wǎng)間孔隙增大,三維網(wǎng)狀結(jié)構(gòu)逐漸不規(guī)則成形,膜降解為碎片,最終觀察不到明顯的網(wǎng)狀結(jié)構(gòu)。(4)蘇木精-伊紅染色觀察:四組鏡下均表現(xiàn)為紅染的纖維蛋白條索逐漸減少,可見炎性細(xì)胞的浸潤,成纖維細(xì)胞和新生毛細(xì)血管數(shù)量逐漸增多。術(shù)后第3d、5d、7d,A1組炎性反應(yīng)都較A2組重(P0.05),B1組炎性反應(yīng)也較B2組重(P0.05);術(shù)后10d,A1與A2間及B1與B2組間炎性反應(yīng)無統(tǒng)計學(xué)差異(P0.05)。結(jié)論:1.A-PRF與PRF膜均可在人工唾液中發(fā)生降解,PRF膜的完全降解時間為14天,A-PRF膜為17天,PRF膜在人工口腔環(huán)境中的降解速度比A-PRF膜快。2.在兔口腔內(nèi),暴露條件加快了膜的降解。3.在兔口腔埋置條件下,A-PRF膜的降解速度慢于PRF膜。4.A-PRF和PRF膜不可單獨(dú)作為屏障膜使用。5.A-PRF和PRF膜的降解為纖維蛋白的降解和三維網(wǎng)狀結(jié)構(gòu)的崩塌。6.A-PRF和PRF膜在暴露條件下會加重炎癥反應(yīng)。
[Abstract]:OBJECTIVE: To compare and analyze the degradation characteristics of platelet-rich fibrin (PRF) and modified platelet-rich fibrin (A-PRF) in vitro and in vivo, and to evaluate their clinical significance. METHODS: 1. In vitro degradation experiment: 10 volunteers were used to prepare A-PRF and PRF membranes from elbow vein blood. A-PRF was prepared by Ghanaati method (1500rpm, 14min), PRF by Choukroun method (2700rpm, 12min). The prepared A-PRF and PRF membranes were immersed in artificial saliva. 36 rabbits were randomly divided into A-PRF group (group A) and PRF group (group B). Each rabbit took 10 ml of blood to prepare A-PRF or PRF membranes, A-PRF (2000 rpm, 24 min), PRF (3250 rpm, 10 min). The prepared membranes were trimmed into 6 mm diameters. Two models were made at each rabbit's hard palate: a 5 mm diameter soft tissue defect was made at the middle line of the palate 4.5 mm from the maxillary medial incisor, the mucoperiosteal was stripped to expose the bone surface, and the soft tissue defect was separated laterally for 1 mm; a 5 mm long corner incision was made at a palatal fold 2 mm below the soft tissue defect, and the whole layer was turned over. In group A, autologous A-PRF circular diaphragm was implanted in the soft tissue defect area, and the edge was sutured and fixed (group A1, exposed group of A-PRF membrane); in group A2, exposed group of A-PRF circular diaphragm was implanted and sutured (group A2, implanted group of A-PRF membrane). In group B, autologous PRF circular diaphragm was implanted in two places, and in group B1, exposed group of PRF membrane in the soft tissue defect area. Zone B2 (PRF membrane embedding group). Each of the four groups had 18 experimental samples. Three animals were randomly killed in three days, five days, seven days, ten days, fourteen days, twenty-one days, A and B after operation. The membrane specimens in the upper jaw were taken out for morphological observation and weighing to calculate the degradation rate. The degradation process was observed by scanning electron microscopy, HE staining, histopathological changes were observed and inflammatory scores were made. In vitro degradation test: (1) All PRF membrane specimens were completely degraded on the 14th day and all A-PRF membrane specimens were completely degraded on the 17th day in artificial saliva culture environment. (2) The degradation rate of A-PRF and PFF membrane was different (P 0.05). 2. In rabbit oral implantation test: (1) Morphological observation: wound healing was observed in groups A2 and B2 on the 7th day after operation; wounds in groups A1 and B1 on the 14th day after operation. On the 7th day after operation, the membrane material in group B1 was obviously absorbed and difficult to be completely removed; on the 10th day after operation, there was no visible specimen in group B1, and it was difficult to remove membrane in group A1 and B2; on the 14th day after operation, it was difficult to remove membrane in group A2, and there was no visible specimen in group A1 and B2; on the 21st day after operation, no implanted specimen could be found in group A2. (2) Degradation: The time of complete degradation was 14 days in group A1, 21 days in group A2 and B1. The degradation rates of membrane samples in A1 and A2 groups at the first five time points were statistically different (P 0.05); the degradation rates of B1 and B2 groups were not statistically different except the third day (P = 0.0660.05), the other time points were statistically significant (P 0.05); A1 and B1 groups on the fifth, seventh and tenth day (P 0.05); A2 and B2 groups on the fifth, seventh and seventh day (P 0.05); Scanning electron microscopy (SEM) results showed that both A-PRF and PRF membranes had three-dimensional network structure, which was connected by trigeminal structure. (4) Hematoxylin-eosin staining showed that the number of red-stained fibrin bands decreased gradually, inflammatory cells infiltrated, fibroblasts and new capillaries increased gradually. Inflammatory reactions in group A1 were heavier than those in group A2 (P 0.05) and group B1 (P 0.05) on the 3rd, 5th, 7th day after operation. There was no significant difference in inflammatory reaction between A1 and A2 and between B1 and B2 groups (P 0.05). Conclusion: 1. Both A-PRF and PRF membranes can be degraded in artificial saliva. The complete degradation time of PRF membranes is 14 days, A-PRF membranes are 17 days, and PRF membranes are degraded faster in artificial oral environment than A-PRF membranes. Dew accelerated the degradation of the membrane. 3. The degradation rate of A-PRF membrane was slower than that of PRF membrane. 4. A-PRF and PRF membrane could not be used as barrier membrane alone. 5. A-PRF and PRF membrane could degrade into fibrin degradation and collapse of three-dimensional network structure.
【學(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R783.6
本文編號：2202430
[Abstract]:OBJECTIVE: To compare and analyze the degradation characteristics of platelet-rich fibrin (PRF) and modified platelet-rich fibrin (A-PRF) in vitro and in vivo, and to evaluate their clinical significance. METHODS: 1. In vitro degradation experiment: 10 volunteers were used to prepare A-PRF and PRF membranes from elbow vein blood. A-PRF was prepared by Ghanaati method (1500rpm, 14min), PRF by Choukroun method (2700rpm, 12min). The prepared A-PRF and PRF membranes were immersed in artificial saliva. 36 rabbits were randomly divided into A-PRF group (group A) and PRF group (group B). Each rabbit took 10 ml of blood to prepare A-PRF or PRF membranes, A-PRF (2000 rpm, 24 min), PRF (3250 rpm, 10 min). The prepared membranes were trimmed into 6 mm diameters. Two models were made at each rabbit's hard palate: a 5 mm diameter soft tissue defect was made at the middle line of the palate 4.5 mm from the maxillary medial incisor, the mucoperiosteal was stripped to expose the bone surface, and the soft tissue defect was separated laterally for 1 mm; a 5 mm long corner incision was made at a palatal fold 2 mm below the soft tissue defect, and the whole layer was turned over. In group A, autologous A-PRF circular diaphragm was implanted in the soft tissue defect area, and the edge was sutured and fixed (group A1, exposed group of A-PRF membrane); in group A2, exposed group of A-PRF circular diaphragm was implanted and sutured (group A2, implanted group of A-PRF membrane). In group B, autologous PRF circular diaphragm was implanted in two places, and in group B1, exposed group of PRF membrane in the soft tissue defect area. Zone B2 (PRF membrane embedding group). Each of the four groups had 18 experimental samples. Three animals were randomly killed in three days, five days, seven days, ten days, fourteen days, twenty-one days, A and B after operation. The membrane specimens in the upper jaw were taken out for morphological observation and weighing to calculate the degradation rate. The degradation process was observed by scanning electron microscopy, HE staining, histopathological changes were observed and inflammatory scores were made. In vitro degradation test: (1) All PRF membrane specimens were completely degraded on the 14th day and all A-PRF membrane specimens were completely degraded on the 17th day in artificial saliva culture environment. (2) The degradation rate of A-PRF and PFF membrane was different (P 0.05). 2. In rabbit oral implantation test: (1) Morphological observation: wound healing was observed in groups A2 and B2 on the 7th day after operation; wounds in groups A1 and B1 on the 14th day after operation. On the 7th day after operation, the membrane material in group B1 was obviously absorbed and difficult to be completely removed; on the 10th day after operation, there was no visible specimen in group B1, and it was difficult to remove membrane in group A1 and B2; on the 14th day after operation, it was difficult to remove membrane in group A2, and there was no visible specimen in group A1 and B2; on the 21st day after operation, no implanted specimen could be found in group A2. (2) Degradation: The time of complete degradation was 14 days in group A1, 21 days in group A2 and B1. The degradation rates of membrane samples in A1 and A2 groups at the first five time points were statistically different (P 0.05); the degradation rates of B1 and B2 groups were not statistically different except the third day (P = 0.0660.05), the other time points were statistically significant (P 0.05); A1 and B1 groups on the fifth, seventh and tenth day (P 0.05); A2 and B2 groups on the fifth, seventh and seventh day (P 0.05); Scanning electron microscopy (SEM) results showed that both A-PRF and PRF membranes had three-dimensional network structure, which was connected by trigeminal structure. (4) Hematoxylin-eosin staining showed that the number of red-stained fibrin bands decreased gradually, inflammatory cells infiltrated, fibroblasts and new capillaries increased gradually. Inflammatory reactions in group A1 were heavier than those in group A2 (P 0.05) and group B1 (P 0.05) on the 3rd, 5th, 7th day after operation. There was no significant difference in inflammatory reaction between A1 and A2 and between B1 and B2 groups (P 0.05). Conclusion: 1. Both A-PRF and PRF membranes can be degraded in artificial saliva. The complete degradation time of PRF membranes is 14 days, A-PRF membranes are 17 days, and PRF membranes are degraded faster in artificial oral environment than A-PRF membranes. Dew accelerated the degradation of the membrane. 3. The degradation rate of A-PRF membrane was slower than that of PRF membrane. 4. A-PRF and PRF membrane could not be used as barrier membrane alone. 5. A-PRF and PRF membrane could degrade into fibrin degradation and collapse of three-dimensional network structure.
【學(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R783.6
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