The Effects of Link N and Bone Marrow-Derived Mesenchymal Stem Cells on the Regeneration of Intervertebral Disc
Bibliographic record
Abstract
Introduction Link N is a naturally occurring peptide that can stimulate regeneration of the intervertebral disc (IVD). IVD repair can also potentially be enhanced by mesenchymal stem cell (MSC) supplementation to maximize extracellular matrix production. In a previous study, we have shown that Link N can inhibit osteogenesis and increase the chondrogenesis of MSCs in vitro. The aim of the present study is to determine the effects of injecting MSCs and Link N alone or together on the repair of degenerated discs. Materials and Methods The largest first four caudal discs with intact cartilaginous endplates were isolated from the tails of 24- to 30-month-old bovines. After a 3-day preconditioning period, the discs were injected with 100 μg trypsin in 75 μL PBS, to induce degeneration. They were then cultured for another 4 days, followed by the injection of MSCs (10 5 ), Link N (75 μg), or a combination of MSCs and Link N, in a final volume of 75 μL PBS. Seven of the trypsin-treated discs were injected with PBS alone to serve as a degeneration control. After the discs were cultured for another 14 days, the extracellular matrix (ECM) proteins and proteoglycans were extracted from the inner nucleus pulposus of the discs by guanidinium chloride. Sulfated glycosaminoglycans (GAGs) were analyzed in tissue extracts ( n = 7/group) by the dimethyl methylene blue (DMMB) dye-binding assay. The expression of type II collagen was analyzed by Western blot. To track the MSCs after injection, MSCs were labeled with PKH67 and observed under confocal microscopy when the discs were dissected. Results After injection, MSCs were distributed throughout the discs and retained their morphology. When the trypsin-treated discs were injected with PBS only, the final GAG concentration in the ECM was 15.12 µg/mg wet weight tissue. When the degenerated discs were injected with MSCs only, the GAG concentration increased to 25.66 µg/mg wet weight tissue. When the degenerated discs were injected with Link N alone or both Link N and MSCs, the GAG concentrations increased to 29.63 and 32.66 µg/mg wet weight tissue, respectively. Compared with the degeneration control, the expression of type II collagen also increased when the degenerated discs were injected with MSCs and Link N, either alone or together. Conclusion MSCs and Link N can both increase GAG production in degenerated discs, with Link N showing a greater effect than MSCs when administered separately. However, an additive effect was found when MSCs and Link N were administered together. The injection of MSCs and Link N can also increase the expression of type II collagen in the degenerated discs. Thus, the administration of MSCs and Link N has therapeutic potential in intervertebral disc repair. Disclosure of Interest None declared
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.001 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.001 | 0.000 |
| Insufficient payload (model declined to judge) | 0.001 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".