Biological properties, degradation and absorption of collagen sponges in vivo

doi:10.3969/j.issn.2095-4344.2014.34.018

Abstract

Abstract:

BACKGROUND: Collagen sponges are applied for hemostatic use, wound healing, and residual cavity filling, which have great values in clinical application and scientific research.

OBJECTIVE: To investigate the biological properties, biocompatibility and biodegradability of collagen sponges in vivo.

METHODS: The spatial structure, pore diameter and porosity of collagen sponges were characterized by scanning electron microscopy. Transmission electron microscopy was used to observe the conformation of collagen sponges. The secondary structure and thermal denaturation temperature of collagen sponges were analyzed by circular dichroism spectrum. Collagen sponges were implanted intramuscularly into the spinal cord of New Zealand rabbits to observe the degradation and absorption and histological changes in vivo.

RESULTS AND CONCLUSION: Collagen sponges had porous structure with varying pore sizes ranging 40-150 μm, the mean pore size of 100 μm, the thickness wall of 1 μm, and a porosity of approximately 95.8%. Collagen sponges had a typical porous structure and periodic light and dark zones. The solution of collagen sponges had a weak positive band near 220 nm and an intense negative band near 206 nm, which indicated a classic triple helix. And the secondary structure and thermal stability of collagen sponges were similar to that of liquid collagen. Collagen sponges began to degrade at 4 weeks, and remained 20% at 12 weeks. These sponges had been associated with foreign body response and inflammation within 2 weeks after implantation. With wound healing, inflammatory reactions gradually reduced and disappeared. During the implantation and degradation of sponges, no significant fibrous capsule formed and no tissue necrosis occurred at implantation site, indicating that collagen sponges have good performance in bioactivity, biocompatibility and degradation.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

全文链接：

Key words: collagen, egg white, materials testing

CLC Number:

R318

Chi Yan-yan, Le Yao-jin, Liu Xu-zhao, Li Qi, Lei Jing, Tang Shun-qing. Biological properties, degradation and absorption of collagen sponges in vivo[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(34): 5515-5519.

Figures/Tables 1

2.1 实验动物数量分析 24只新西兰兔均进入结果分析。 2.2 胶原蛋白海绵的电镜分析扫描电镜分析经测定胶原蛋白海绵孔径分布为40-150 μm，平均100 μm，孔壁 1 μm左右，孔隙率约95.8%，呈纤维细丝网状结构，见图1A。图1B显示的是胶原蛋白海绵的透射电镜图，胶原纤维呈现分散态，纤维直径200-300 nm，可观察到胶原具有的特殊的周期性横纹区带。 2.3 胶原蛋白海绵的圆二色谱分析胶原由3条α肽链组成，α肽链自身为α螺旋结构，3条α肽链则以平行、右手螺旋形式缠绕成“草绳状”三股螺旋结构。采用圆二色谱法测定胶原溶液、胶原蛋白海绵二级结构特征，结果显示两者图谱具有非常好的重叠性(图1C)，均在220 nm有一强的正特征吸收峰，206 nm左右有一强负特征吸收峰，具有典型胶原蛋白三螺旋结构的特征吸收性质，说明在胶原溶液冻干成海绵过程中胶原构象并未发生变化。对圆二色谱所测数据采用Cdpro软件分析胶原蛋白海绵中不同二级结构所占的比例，其中α螺旋含量为34.4%，β折叠为33%，β转角为16.8%，无规卷曲为16.4%。与对照组胶原溶液一致。圆二色谱同时可以反映胶原溶液与胶原蛋白海绵的变性温度，当胶原蛋白的三螺旋结构被完全破坏时，正峰会完全消失。如图2所示，以220 nm处吸收峰的变化为例说明，纵坐标反映二级结构α螺旋构象的变化情况，显示胶原溶液在33 ℃之后正吸收峰值开始降低，构象遭到破坏，随着温度进一步升高，当温度达到50 ℃左右，蛋白变性达到最大程度；胶原蛋白海绵在30 ℃左右正吸收峰值开始降低，随着温度升高到达48 ℃构象完全遭到破坏，二者变性温度基本一致。 2.4 胶原蛋白海绵动物体内降解吸收的组织病理观察实验组术后2周伤口痊愈，未出现出血、红肿、坏死和试样排出等异常现象，海绵的降解率为0。术后第2，4，8，12周取材，第4周可见到胶原蛋白海绵，海绵的降解率为0，第8周海绵的降解率为33%，至12周海绵降解率达75%(图3)。病理组织切片分析显示，第2周植入部位有明显异物反应，可见较多单核细胞及少量异物巨细胞浸润，组织炎症反应小于Ⅲ级(图4A)；第4周植入部位可见较多增生结缔组织，其中散在单核细胞浸润，未见异物巨细胞，炎症反应小于Ⅰ级，且未见明显纤维囊结构(图4B)；术后第8周，植入部位可见较多纤维结缔组织增生并发生玻璃样变，散在单核细胞浸润，未见异物巨细胞(图4C)；随着植入时间达12周，植入部位未见单核细胞、淋巴细胞及异物巨细胞浸润，炎症反应逐渐消失，可见脂肪细胞，且有结缔组织增生(图4D)。空白对照组在实验过程中均未出现异物反应(图4E)。"

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