骨損傷修復中H型血管的分布和作用及血管束神經束聯(lián)合植入組織工程骨修復羊大段脛骨缺損的研究
發(fā)布時間：2018-12-17 03:00
【摘要】：骨組織是一個高度血管化和神經化的組織。無論在正常的骨損傷修復過程中還是在應用組織工程骨修復骨缺損的情況下,及時和豐富的血管再生是保證骨損傷正常修復和組織工程骨存活的基礎。最近有文獻報道在小鼠骨發(fā)育過程中存在兩種血管內皮細胞亞型,即高表達CD31和Emcn的H型內皮細胞和低表達CD31和Emcn的L型內皮細胞。這兩種內皮細胞亞型具有不同的形態(tài)、分布和功能特征。其中H型內皮細胞在介導小鼠骨發(fā)育成血管和成骨偶聯(lián)的過程中起著重要作用。本研究第一部分對H型血管在正常成年大鼠長骨中的分布以及在正常和骨質疏松大鼠骨損傷修復中的分布和變化規(guī)律進行研究,目的是對H型血管在骨損傷修復中的地位和作用獲得更深入的認識。通過研究我們發(fā)現(xiàn),在正常成年大鼠長骨中也存在H型血管并特異的分布在靠近干骺端和骨內膜的骨小梁表面這些成骨活動活躍的部位;在正常大鼠骨損傷的修復過程中,H型血管在損傷后第3d即出現(xiàn)大量增生和分布;隨著修復時間的進展,H型血管的量逐漸減少,但特異性的分布在靠近生長前沿的新生骨小梁的表面,在遠離生長前沿的骨小梁表面分布顯著減少;作為對照,在骨質疏松大鼠骨損傷修復的各個時期,H型血管增生的量均顯著減少,并且未發(fā)現(xiàn)其特異性高分布,新生骨小梁數(shù)量和厚度顯著減少且結構紊亂。結合以往的文獻推測H型血管具有支持大鼠骨發(fā)育和骨損傷修復中新生骨小梁生長的作用,骨質疏松大鼠骨損傷修復中H型血管數(shù)量的顯著減少是其成骨量少的重要原因。此外,我們還研究了在正常和骨質疏松大鼠骨損傷修復中成血管相關因子Hif-1α、VEGFA和成骨因子BMP2、Osterix轉錄水平的變化。研究發(fā)現(xiàn)在骨損傷后第3d,骨質疏松組Hif-1α和VEGFA的轉錄水平顯著高于正常組;但在術后第7d和第14d,骨質疏松組Hif-1α和VEGFA的轉錄水平顯著低于正常組;骨質疏松組成骨因子BMP2和Osterix的轉錄水平在術后3d、7d和14d均顯著低于正常組;研究提示骨質疏松大鼠在損傷修復期成血管和成骨因子轉錄水平的顯著降低也是其成血管和成骨量低下的原因。組織工程骨血管化和神經化程度不足是制約其向臨床應用轉化的重要原因。有學者發(fā)現(xiàn)將動靜脈環(huán)或動靜脈束移植入組織工程骨來修復骨缺損可以顯著促進組織工程骨血管化和骨缺損的修復。另有學者發(fā)現(xiàn)將感覺神經束離斷后移植入組織工程骨中也能起到與血管束植入相同的促成骨效果,其中的原因可能是移植入的神經束本身具有滋養(yǎng)血管,而且感覺神經束的植入可以通過CGRP、NPY和SP等外分泌遞質的釋放促進種子細胞的存活和成骨。據(jù)此,裴國獻教授提出了組織工程骨血管化和神經化同步構建的理論。本研究第二部分通過分別將隱血管束與隱神經束同時植入組織工程骨、單純隱血管束植入組織工程骨和單純應用組織工程骨修復綿羊脛骨3cm大節(jié)段骨缺損,在術后3個月分別通過X線平片、微米X射線掃描后三維重建、生物力學檢測以及VG染色評價各組骨缺損的修復效果和生物力學功能。結果顯示,隱血管束和隱神經束聯(lián)合植入組成骨量和材料的降解率均顯著高于其他兩組,并且獲得了較其他兩組更好的生物力學功能的恢復。本研究的結果提示組織工程骨血管化和神經化同步構建對于促進組織工程骨成骨和力學功能的恢復具有重要作用。
[Abstract]:Bone tissue is a highly vascularized and neurotic tissue. In the case of the normal bone injury repair or the application of the tissue engineering bone to repair the bone defect, the timely and abundant blood vessel regeneration is the basis for ensuring the normal repair of the bone injury and the survival of the tissue engineering bone. Recently, there are two types of vascular endothelial cell subtypes, namely, H-type endothelial cells with high expression of CD31 and Emcn, and L-type endothelial cells with low expression of CD31 and Emcn in the development of mouse bone. The two types of endothelial cells have different morphology, distribution and functional characteristics. in which H-type endothelial cells play an important role in mediating the development of a mouse bone into a blood vessel and a bone-forming coupling. The first part of this study is to study the distribution of H-type blood vessels in the long bones of normal adult rats and the distribution and the rule of change in the repair of bone injury in normal and osteoporotic rats. The purpose of this study is to get a more thorough understanding of the status and role of H-type blood vessels in the repair of bone injury. We have found that H-type blood vessels are also present in the long bones of the normal adult rats and are specifically distributed on the surface of the bone small beams close to the dry end and the inner surface of the bone, which are active parts of the bone; during the repair of the bone injury of the normal rats, In the third day after the injury, the H-type blood vessel has a large number of hyperplasia and distribution; with the development of the repair time, the amount of the H-type blood vessel is gradually reduced, but the specific distribution is close to the surface of the new-born bone small beam near the growth front, and the surface distribution of the small-beam surface which is far from the leading edge of the growth is significantly reduced; As a control, the amount of H-type vessel hyperplasia was significantly reduced in the various stages of bone injury repair in the osteoporosis rats, and no specific high profile was found, with a significant reduction in the number and thickness of the new bone and a structural disorder. Combined with the previous literature, the H-type blood vessel has the effect of supporting the growth of the new bone in the bone development and the bone injury repair of the rat, and the significant reduction of the number of H-type blood vessels in the bone injury repair of the osteoporosis rats is an important cause of the small amount of the bone. In addition, we have also studied the changes of vascular-related factors Hif-1, BFA and bone factor BMP2 and Osternix in the repair of bone injury in normal and osteoporotic rats. The results showed that the levels of Hif-1 and HFA in the osteoporosis group were significantly higher than those in the normal group after the bone injury, but the levels of Hif-1 and HFA in the osteoporosis group were significantly lower than that of the normal group at the 7th and 14th day after the operation, and the transcription level of the bone factors BMP2 and Osterx in the osteoporosis group was lower than that of the normal group. Both 7d and 14d were significantly lower than that in the normal group. The insufficient degree of vascularization and neurotization of the tissue engineering is an important factor in the transformation of the clinical application. It is found that the graft of the arteriovenous ring or the arteriovenous beam into the tissue engineering bone to repair the bone defect can significantly promote the tissue engineering bone vascularization and the repair of the bone defect. another scholar has found that it is also possible to implant the sensory nerve bundle into the tissue engineered bone after it is broken, which may be the same contributing bone effect as the implantation of the vascular bundle, where the cause may be that the implanted nerve bundle itself has a nourishing blood vessel, and the implantation of the sensory nerve bundle may be through the cgrp, The release of exogenous hormones, such as NPY and SP, promotes the survival of seed cells and the formation of bone. On the basis of this, the author of the founding of the Republic of China put forward the theory of the construction of the tissue engineering's vascularisation and the synchronization of the neurogenesis. in that second part of the study, the bone defect of the 3cm large section of the sheep's tibia was repaired with the bone of the tissue engineer and the saphenous nerve bundle at the same time as the saphenous bundle and the saphenous nerve bundle, and the bone defect of the 3cm large section of the sheep tibia was repaired by using the tissue engineering bone and the pure application tissue engineering bone, respectively, and the bone defect of the bone of the sheep tibia is repaired through the X-ray flat sheet at 3 months after the operation, Three-dimensional reconstruction, biomechanical testing and VG staining were used to evaluate the repair and biomechanical function of each group of bone defects. The results showed that the rate of bone mass and material degradation was significantly higher than that of other two groups, and the recovery of other two groups of better biomechanical function was obtained. The results of this study suggest that the construction of the tissue engineering bone and the synchronization of the neurogenesis play an important role in promoting the recovery of bone and mechanical functions of the tissue engineering.
【學位授予單位】：第四軍醫(yī)大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：R68
，
本文編號：2383536
[Abstract]:Bone tissue is a highly vascularized and neurotic tissue. In the case of the normal bone injury repair or the application of the tissue engineering bone to repair the bone defect, the timely and abundant blood vessel regeneration is the basis for ensuring the normal repair of the bone injury and the survival of the tissue engineering bone. Recently, there are two types of vascular endothelial cell subtypes, namely, H-type endothelial cells with high expression of CD31 and Emcn, and L-type endothelial cells with low expression of CD31 and Emcn in the development of mouse bone. The two types of endothelial cells have different morphology, distribution and functional characteristics. in which H-type endothelial cells play an important role in mediating the development of a mouse bone into a blood vessel and a bone-forming coupling. The first part of this study is to study the distribution of H-type blood vessels in the long bones of normal adult rats and the distribution and the rule of change in the repair of bone injury in normal and osteoporotic rats. The purpose of this study is to get a more thorough understanding of the status and role of H-type blood vessels in the repair of bone injury. We have found that H-type blood vessels are also present in the long bones of the normal adult rats and are specifically distributed on the surface of the bone small beams close to the dry end and the inner surface of the bone, which are active parts of the bone; during the repair of the bone injury of the normal rats, In the third day after the injury, the H-type blood vessel has a large number of hyperplasia and distribution; with the development of the repair time, the amount of the H-type blood vessel is gradually reduced, but the specific distribution is close to the surface of the new-born bone small beam near the growth front, and the surface distribution of the small-beam surface which is far from the leading edge of the growth is significantly reduced; As a control, the amount of H-type vessel hyperplasia was significantly reduced in the various stages of bone injury repair in the osteoporosis rats, and no specific high profile was found, with a significant reduction in the number and thickness of the new bone and a structural disorder. Combined with the previous literature, the H-type blood vessel has the effect of supporting the growth of the new bone in the bone development and the bone injury repair of the rat, and the significant reduction of the number of H-type blood vessels in the bone injury repair of the osteoporosis rats is an important cause of the small amount of the bone. In addition, we have also studied the changes of vascular-related factors Hif-1, BFA and bone factor BMP2 and Osternix in the repair of bone injury in normal and osteoporotic rats. The results showed that the levels of Hif-1 and HFA in the osteoporosis group were significantly higher than those in the normal group after the bone injury, but the levels of Hif-1 and HFA in the osteoporosis group were significantly lower than that of the normal group at the 7th and 14th day after the operation, and the transcription level of the bone factors BMP2 and Osterx in the osteoporosis group was lower than that of the normal group. Both 7d and 14d were significantly lower than that in the normal group. The insufficient degree of vascularization and neurotization of the tissue engineering is an important factor in the transformation of the clinical application. It is found that the graft of the arteriovenous ring or the arteriovenous beam into the tissue engineering bone to repair the bone defect can significantly promote the tissue engineering bone vascularization and the repair of the bone defect. another scholar has found that it is also possible to implant the sensory nerve bundle into the tissue engineered bone after it is broken, which may be the same contributing bone effect as the implantation of the vascular bundle, where the cause may be that the implanted nerve bundle itself has a nourishing blood vessel, and the implantation of the sensory nerve bundle may be through the cgrp, The release of exogenous hormones, such as NPY and SP, promotes the survival of seed cells and the formation of bone. On the basis of this, the author of the founding of the Republic of China put forward the theory of the construction of the tissue engineering's vascularisation and the synchronization of the neurogenesis. in that second part of the study, the bone defect of the 3cm large section of the sheep's tibia was repaired with the bone of the tissue engineer and the saphenous nerve bundle at the same time as the saphenous bundle and the saphenous nerve bundle, and the bone defect of the 3cm large section of the sheep tibia was repaired by using the tissue engineering bone and the pure application tissue engineering bone, respectively, and the bone defect of the bone of the sheep tibia is repaired through the X-ray flat sheet at 3 months after the operation, Three-dimensional reconstruction, biomechanical testing and VG staining were used to evaluate the repair and biomechanical function of each group of bone defects. The results showed that the rate of bone mass and material degradation was significantly higher than that of other two groups, and the recovery of other two groups of better biomechanical function was obtained. The results of this study suggest that the construction of the tissue engineering bone and the synchronization of the neurogenesis play an important role in promoting the recovery of bone and mechanical functions of the tissue engineering.
【學位授予單位】：第四軍醫(yī)大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：R68
，
本文編號：2383536
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