The chitosan collagen salvianolic membrane mitigates intervertebral disc degeneration through suppression of the Wnt/β-catenin signaling.

IF 2 4区 生物学 Q4 CELL BIOLOGY
Zhaozhong Yu, Jun Kuang, Xuan Wan, Chaoyi Liang, Fan Wu, Zhimin Shu, Weidong Liang, Xiaoming Wan, Jianbin Gong
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引用次数: 0

Abstract

Intervertebral disc degeneration (IVDD), a natural aging process, leads to intervertebral disc pain, with lumbar resection providing pain relief but potential postoperative complications affecting outcomes. The chitosan-collagen-salvianolic (CCS) membrane has shown promise in reducing scar formation after lumbar resection, though the mechanisms remain unclear. A lumbar laminectomy model was used, with materials applied to the surgical site. Tissue samples were collected at 4 and 8 wk post-surgery for histological evaluation to assess pathological changes and apoptosis. Western blot analysis examined protein expression in the Wnt/β-catenin signaling pathway in scar tissue. Rabbit vascular adventitia fibroblast (VAF) cells were isolated, and a Wnt3a overexpression vector was created. Various composite membrane materials and pathway inhibitors were tested, with assessments of cell invasion, apoptosis, viability, cytokine levels, and protein expression. Results showed that lumbar laminectomy activated the Wnt/β-catenin pathway. The CCS membrane facilitated healing by inhibiting this pathway. It enhanced VAF cell activity, reduced apoptosis, and suppressed Wnt3a pathway activity. Additionally, it protected cells from Wnt3a overexpression-induced damage while maintaining normal function. In conclusion, the CCS composite membrane promoted bone tissue repair after lumbar laminectomy by inhibiting Wnt/β-catenin pathway activation and preserving extracellular matrix fibroblast function.

壳聚糖胶原丹酚膜通过抑制Wnt/β-catenin信号通路减轻椎间盘退变。
椎间盘退变(IVDD)是一种自然老化过程,会导致椎间盘疼痛,腰椎切除术可以缓解疼痛,但潜在的术后并发症会影响结果。壳聚糖-胶原-丹酚(CCS)膜在腰椎切除术后减少疤痕形成方面显示出希望,尽管机制尚不清楚。采用腰椎椎板切除术模型,将材料应用于手术部位。分别于术后4周和8周采集组织标本进行组织学评估,以评估病理变化和细胞凋亡情况。Western blot检测瘢痕组织中Wnt/β-catenin信号通路的蛋白表达。分离兔血管外膜成纤维细胞(VAF),构建Wnt3a过表达载体。测试了各种复合膜材料和途径抑制剂,评估细胞侵袭、凋亡、活力、细胞因子水平和蛋白质表达。结果显示,腰椎椎板切除术激活了Wnt/β-catenin通路。CCS膜通过抑制这一途径促进愈合。增强VAF细胞活性,减少凋亡,抑制Wnt3a通路活性。此外,它可以保护细胞免受Wnt3a过表达诱导的损伤,同时保持正常功能。综上所述,CCS复合膜通过抑制Wnt/β-catenin通路激活和保持细胞外基质成纤维细胞功能,促进腰椎椎板切除术后骨组织修复。
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来源期刊
CiteScore
3.70
自引率
4.80%
发文量
96
审稿时长
3 months
期刊介绍: In Vitro Cellular & Developmental Biology - Animal is a journal of the Society for In Vitro Biology (SIVB). Original manuscripts reporting results of research in cellular, molecular, and developmental biology that employ or are relevant to organs, tissue, tumors, and cells in vitro will be considered for publication. Topics covered include: Biotechnology; Cell and Tissue Models; Cell Growth/Differentiation/Apoptosis; Cellular Pathology/Virology; Cytokines/Growth Factors/Adhesion Factors; Establishment of Cell Lines; Signal Transduction; Stem Cells; Toxicology/Chemical Carcinogenesis; Product Applications.
文献相关原料
原料信息
原料厂家
1
anti-Wnt3a
2
flow cytometer
3
RIPA lysis buffer
4
RIPA lysis buffer
5
ELISA reader
6
anti-Bcl-2
7
vimentin antibody
8
CFX Connect
9
highly sensitive chemiluminescence imaging system
10
anti-Caspase3
11
ultrapure RNA extraction kit
12
BCA protein quantification kit
13
sodium hyaluronate
14
vacuum freeze dryer
15
New Zealand white rabbit
16
CCK-8 reagent
18
PVDF membrane
19
Annexin V-FITC
20
UV-visible spectrophotometer
21
CKX53 fluorescence microscope
22
fluorescence microscope
23
microscope
24
anti-Bax
25
anti-GSK-3β
26
anti-HDAC1
27
anti-p-GSK3β (Tyr216)
28
anti-TGF-β1
29
anti-TNF-α
30
anti-β-catenin
31
secondary antibody IgG (H+L)
32
magnetic stirrer
33
anti-GAPDH
34
RNA reverse transcription kit
35
hematoxylin
36
22
chitosan
9012-76-4
¥24.00-¥24532.00
37
22
crystal violet
548-62-9
¥19.00-¥26733.86
38
22
salvianolic
76822-21-4
¥36.00-¥15763.00
39
collagen type I
9007-34-5
¥30.00-¥50800.56
40
22
eosin
17372-87-1
¥20.00-¥6009.00
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