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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Pathology, Diagnostics, and Therapeutics".

Deadline for manuscript submissions: closed (31 March 2023) | Viewed by 27941

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Prof. Dr. Inbo Han

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Department of Neurosurgery, CHA University School of Medicine, CHA Bundang Medical Center, Seongnam-si 13496, Republic of Korea
Interests: spinal diseases (spinal cord injury, intervertebral disc degeneration, osteoporosis, etc.); pain; regeneration; stem cell; neuroprotection
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Dr. Takashi Yurube

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Department of Orthopaedic Surgery, Kobe University Graduate School of Medicine, Kobe 650-0017, Japan
Interests: minimally invasive reconstruction; spine surgery; regenerative medicine
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Dr. Daisuke Sakai

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Guest Editor

Department Orthopaedic Surgery, Tokai University School of Medicine, 143 Shimokasuya, Isehara 259-1193, Japan
Interests: intervertebral disc regeneration
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

It is my pleasure to inform you about the Special Issue “Regeneration for Spinal Diseases”. Spinal diseases place a significant burden on the general population. The prevalence of degenerative spinal diseases, including intervertebral disc degeneration and osteoporosis, increases with age, and the affected population may suffer from long-term disability. The most serious conditions affecting the spine include spinal cord injury due to traumatic or non-traumatic causes (e.g., cancer, infection). Despite recent advancements in the management of these spinal diseases, there is growing research interest in discovering a novel therapeutic strategy.

This Special Issue focuses on regenerative therapy for spinal diseases, including spinal cord injury, intervertebral disc degeneration, osteoporosis, and pseudarthrosis. It will be published in the International Journal of Molecular Sciences (IJMS, https://www.mdpi.com/journal/ijms, ISSN 1422-0067), and is now open to receive submissions of full research articles and authoritative review papers for peer-review and possible publication.

Due to the success of the 1st and 2nd editions, we would like to add more results and new insights from recent research projects.

https://www.mdpi.com/journal/ijms/special_issues/Spinal_Diseases

https://www.mdpi.com/journal/ijms/special_issues/Spinal_Diseases_2

Prof. Dr. Inbo Han
Dr. Takashi Yurube
Dr. Daisuke Sakai
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. There is an Article Processing Charge (APC) for publication in this open access journal. For details about the APC please see here. Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

regeneration
intervertebral disc degeneration
spinal cord injury
osteoporosis
spinal fusion
muscle regeneration

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Related Special Issues

Regeneration for Spinal Diseases in International Journal of Molecular Sciences (16 articles)
Regeneration for Spinal Diseases 2.0 in International Journal of Molecular Sciences (15 articles)

Published Papers (11 papers)

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Editorial

Jump to: Research, Review

7 pages, 224 KiB

Open AccessEditorial

Concepts of Regeneration for Spinal Diseases in 2023

by Takashi Yurube, Inbo Han and Daisuke Sakai

Int. J. Mol. Sci. 2023, 24(22), 16335; https://doi.org/10.3390/ijms242216335 - 15 Nov 2023

Cited by 2 | Viewed by 1369

Abstract

It is our pleasure to announce the publication of the Special Issue “Regeneration for Spinal Diseases 3.0” in the International Journal of Molecular Sciences (ISSN 1422-0067) [...] Full article

(This article belongs to the Special Issue Regeneration for Spinal Diseases 3.0)

Research

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13 pages, 3738 KiB

Open AccessArticle

Cytokine Inhibitors Upregulate Extracellular Matrix Anabolism of Human Intervertebral Discs under Alginate Beads and Alginate-Embedded Explant Cultures

by Kenichiro Kakutani, Takashi Yurube, Howard S. An, Minoru Doita and Koichi Masuda

Int. J. Mol. Sci. 2023, 24(15), 12336; https://doi.org/10.3390/ijms241512336 - 2 Aug 2023

Cited by 1 | Viewed by 1021

Abstract

We investigated the effects of the cytokine inhibitors IL-1 receptor antagonist (IL-1Ra) and soluble tumor necrosis factor receptor-1 (sTNFR1) on the extracellular matrix metabolism of human intervertebral discs (IVDs) and the roles of IL-1β and TNF in the homeostasis of IVD cells. The 1.2% alginate beads and the explants obtained from 35 human lumbar discs were treated with cytokine inhibitors. Extracellular matrix metabolism was evaluated by proteoglycan (PG) and collagen syntheses and IL-1β, TNF, and IL-6 expressions after three days of culture in the presence or absence of IL-1Ra, sTNFR1, and cycloheximide. Simultaneous treatment with IL-1Ra and sTNFR1 stimulated PG and collagen syntheses in the NP and AF cells and explants. The IL-1β concentration was significantly correlated to the relative increase in PG synthesis in AF explants after simultaneous cytokine inhibitor treatment. The relative increase in PG synthesis induced by simultaneous cytokine treatment was significantly higher in an advanced grade of MRI. Expressions of IL-1β and TNF were upregulated by each cytokine inhibitor, and simultaneous treatment suppressed IL-1β and TNF productions. In conclusion, IL-1Ra and sTNFR1 have the potential to increase PG and collagen synthesis in IVDs. IL-1β and TNF have a feedback pathway to maintain optimal expression, resulting in the control of homeostasis in IVD explants. Full article

(This article belongs to the Special Issue Regeneration for Spinal Diseases 3.0)

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13 pages, 2513 KiB

Open AccessArticle

Investigation of the Mitigation of DMSO-Induced Cytotoxicity by Hyaluronic Acid following Cryopreservation of Human Nucleus Pulposus Cells

by Daiki Munesada, Daisuke Sakai, Yoshihiko Nakamura, Jordy Schol, Erika Matsushita, Shota Tamagawa, Kosuke Sako, Shota Ogasawara, Masato Sato and Masahiko Watanabe

Int. J. Mol. Sci. 2023, 24(15), 12289; https://doi.org/10.3390/ijms241512289 - 31 Jul 2023

Cited by 3 | Viewed by 1683

Abstract

To develop an off-the-shelf therapeutic product for intervertebral disc (IVD) repair using nucleus pulposus cells (NPCs), it is beneficial to mitigate dimethyl sulfoxide (DMSO)-induced cytotoxicity caused by intracellular reactive oxygen species (ROS). Hyaluronic acid (HA) has been shown to protect chondrocytes against ROS. Therefore, we examined the potential of HA on mitigating DMSO-induced cytotoxicity for the enhancement of NPC therapy. Human NPC cryopreserved in DMSO solutions were thawed, mixed with equal amounts of EDTA-PBS (Group E) or HA (Group H), and incubated for 3–5 h. After incubation, DMSO was removed, and the cells were cultured for 5 days. Thereafter, we examined cell viability, cell proliferation rates, Tie2 positivity (a marker of NP progenitor cells), and the estimated numbers of Tie2 positive cells. Fluorescence intensity of DHE and MitoSOX staining, as indicators for oxidative stress, were evaluated by flow cytometry. Group H showed higher rates of cell proliferation and Tie2 expressing cells with a trend toward suppression of oxidative stress compared to Group E. Thus, HA treatment appears to suppress ROS induced by DMSO. These results highlight the ability of HA to maintain NPC functionalities, suggesting that mixing HA at the time of transplantation may be useful in the development of off-the-shelf NPC products. Full article

(This article belongs to the Special Issue Regeneration for Spinal Diseases 3.0)

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23 pages, 6018 KiB

Open AccessArticle

Co-Administration of Resolvin D1 and Peripheral Nerve-Derived Stem Cell Spheroids as a Therapeutic Strategy in a Rat Model of Spinal Cord Injury

by Seung-Young Jeong, Hye-Lan Lee, SungWon Wee, HyeYeong Lee, GwangYong Hwang, SaeYeon Hwang, SolLip Yoon, Young-Il Yang, Inbo Han and Keung-Nyun Kim

Int. J. Mol. Sci. 2023, 24(13), 10971; https://doi.org/10.3390/ijms241310971 - 30 Jun 2023

Cited by 3 | Viewed by 1870

Abstract

Spinal cord injury (SCI), primarily caused by trauma, leads to permanent and lasting loss of motor, sensory, and autonomic functions. Current therapeutic strategies are focused on mitigating secondary injury, a crucial aspect of SCI pathophysiology. Among these strategies, stem cell therapy has shown considerable therapeutic potential. This study builds on our previous work, which demonstrated the functional recovery and neuronal regeneration capabilities of peripheral nerve-derived stem cell (PNSC) spheroids, which are akin to neural crest stem cells, in SCI models. However, the limited anti-inflammatory capacity of PNSC spheroids necessitates a combined therapeutic approach. As a result, we investigated the potential of co-administering resolvin D1 (RvD1), known for its anti-inflammatory and neuroprotective properties, with PNSC spheroids. In vitro analysis confirmed RvD1’s anti-inflammatory activity and its inhibitory effect on pro-inflammatory cytokines. In vivo studies involving a rat SCI model demonstrated that combined therapy of RvD1 and PNSC spheroids outperformed monotherapies, exhibiting enhanced neuronal regeneration and anti-inflammatory effects as validated through behavior tests, quantitative reverse transcription polymerase chain reaction, and immunohistochemistry. Thus, our findings suggest that the combined application of RvD1 and PNSC spheroids may represent a novel therapeutic approach for SCI management. Full article

(This article belongs to the Special Issue Regeneration for Spinal Diseases 3.0)

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21 pages, 5580 KiB

Open AccessArticle

Anti-Inflammatory Effects of Adiponectin Receptor Agonist AdipoRon against Intervertebral Disc Degeneration

by Hiroki Ohnishi, Zhongying Zhang, Takashi Yurube, Yoshiki Takeoka, Yutaro Kanda, Ryu Tsujimoto, Kunihiko Miyazaki, Tomoya Matsuo, Masao Ryu, Naotoshi Kumagai, Kohei Kuroshima, Yoshiaki Hiranaka, Ryosuke Kuroda and Kenichiro Kakutani

Int. J. Mol. Sci. 2023, 24(10), 8566; https://doi.org/10.3390/ijms24108566 - 10 May 2023

Cited by 6 | Viewed by 1994

Abstract

Adiponectin, a hormone secreted by adipocytes, has anti-inflammatory effects and is involved in various physiological and pathological processes such as obesity, inflammatory diseases, and cartilage diseases. However, the function of adiponectin in intervertebral disc (IVD) degeneration is not well understood. This study aimed to elucidate the effects of AdipoRon, an agonist of adiponectin receptor, on human IVD nucleus pulposus (NP) cells, using a three-dimensional in vitro culturing system. This study also aimed to elucidate the effects of AdipoRon on rat tail IVD tissues using an in vivo puncture-induced IVD degeneration model. Analysis using quantitative polymerase chain reaction demonstrated the downregulation of gene expression of proinflammatory and catabolic factors by interleukin (IL)-1β (10 ng/mL) in human IVD NP cells treated with AdipoRon (2 μM). Furthermore, western blotting showed AdipoRon-induced suppression of p65 phosphorylation (p < 0.01) under IL-1β stimulation in the adenosine monophosphate-activated protein kinase (AMPK) pathway. Intradiscal administration of AdipoRon was effective in alleviating the radiologic height loss induced by annular puncture of rat tail IVD, histomorphological degeneration, production of extracellular matrix catabolic factors, and expression of proinflammatory cytokines. Therefore, AdipoRon could be a new therapeutic candidate for alleviating the early stage of IVD degeneration. Full article

(This article belongs to the Special Issue Regeneration for Spinal Diseases 3.0)

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15 pages, 8949 KiB

Open AccessArticle

Human Embryonic Stem-Cell-Derived Exosomes Repress NLRP3 Inflammasome to Alleviate Pyroptosis in Nucleus Pulposus Cells by Transmitting miR-302c

by Yawen Yu, Wenting Li, Tinghui Xian, Mei Tu, Hao Wu and Jiaqing Zhang

Int. J. Mol. Sci. 2023, 24(8), 7664; https://doi.org/10.3390/ijms24087664 - 21 Apr 2023

Cited by 9 | Viewed by 2356

Abstract

Recent studies have shown that the NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3) inflammasome is extensively activated in the process of intervertebral disc degeneration (IVDD), leading to the pyroptosis of nucleus pulposus cells (NPCs) and the exacerbation of the pathological development of the intervertebral disc (IVD). Exosomes derived from human embryonic stem cells (hESCs-exo) have shown great therapeutic potential in degenerative diseases. We hypothesized that hESCs-exo could alleviate IVDD by downregulating NLRP3. We measured the NLRP3 protein levels in different grades of IVDD and the effect of hESCs-exo on the H₂O₂-induced pyroptosis of NPCs. Our results indicate that the expression of NLRP3 was upregulated with the increase in IVD degeneration. hESCs-exo were able to reduce the H₂O₂-mediated pyroptosis of NPCs by downregulating the expression levels of NLRP3 inflammasome-related genes. Bioinformatics software predicted that miR-302c, an embryonic stem-cell-specific RNA, can inhibit NLRP3, thereby alleviating the pyroptosis of NPCs, and this was further verified by the overexpression of miR-302c in NPCs. In vivo experiments confirmed the above results in a rat caudal IVDD model. Our study demonstrates that hESCs-exo could inhibit excessive NPC pyroptosis by downregulating the NLRP3 inflammasome during IVDD, and miR-302c may play a key role in this process. Full article

(This article belongs to the Special Issue Regeneration for Spinal Diseases 3.0)

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15 pages, 5732 KiB

Open AccessArticle

Expression of GADD45G and CAPRIN1 in Human Nucleus Pulposus: Implications for Intervertebral Disc Degeneration

by Koki Kawaguchi, Koji Akeda, Junichi Yamada, Takahiro Hasegawa, Norihiko Takegami, Tatsuhiko Fujiwara and Akihiro Sudo

Int. J. Mol. Sci. 2023, 24(6), 5768; https://doi.org/10.3390/ijms24065768 - 17 Mar 2023

Cited by 3 | Viewed by 2013

Abstract

Marked cellular changes occur in human intervertebral disc (IVD) degeneration during disc degeneration with biochemical changes. Genome-wide analysis of the DNA methylation profile has identified 220 differentially methylated loci associated with human IVD degeneration. Among these, two cell-cycle–associated genes, growth arrest and DNA damage 45 gamma (GADD45G) and cytoplasmic activation/proliferation-associated protein-1 (CAPRIN1), were focused on. The expression of GADD45G and CAPRIN1 in human IVDs remains unknown. We aimed to examine the expression of GADD45G and CAPRIN1 in human nucleus pulposus (NP) cells and evaluate those in human NP tissues in the early and advanced stages of degeneration according to Pfirrmann magnetic resonance imaging (MRI) and histological classifications. Human NP cells were cultured as monolayers after isolation from NP tissues by sequential enzyme digestion. Total RNA was isolated, and the mRNA expression of GADD45G and CAPRIN1 was quantified using real-time polymerase chain reaction. To examine the effects of pro-inflammatory cytokines on mRNA expression, human NP cells were cultured in the presence of IL-1β. Protein expression was evaluated using Western blotting and immunohistochemistry. GADD45G and CAPRIN1 expression was identified in human NP cells at both mRNA and protein levels. The percentage of cells immunopositive for GADD45G and CAPRIN1 significantly increased according to the Pfirrmann grade. A significant correlation between the histological degeneration score and the percentage of GADD45G-immunopositive cells was identified, but not with that of CAPRIN1-immunopositive cells. The expression of cell-cycle-associated proteins (GADD45G and CAPRIN1) was enhanced in human NP cells at an advanced stage of degeneration, suggesting that it may be regulated during the progression of IVD degeneration to maintain the integrity of human NP tissues by controlling cell proliferation and apoptosis under epigenetic alteration. Full article

(This article belongs to the Special Issue Regeneration for Spinal Diseases 3.0)

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21 pages, 8133 KiB

Open AccessArticle

Long-Term SMN- and Ncald-ASO Combinatorial Therapy in SMA Mice and NCALD-ASO Treatment in hiPSC-Derived Motor Neurons Show Protective Effects

by Anixa Muiños-Bühl, Roman Rombo, Karen K. Ling, Eleonora Zilio, Frank Rigo, C. Frank Bennett and Brunhilde Wirth

Int. J. Mol. Sci. 2023, 24(4), 4198; https://doi.org/10.3390/ijms24044198 - 20 Feb 2023

Cited by 4 | Viewed by 2400

Abstract

For SMA patients with only two SMN2 copies, available therapies might be insufficient to counteract lifelong motor neuron (MN) dysfunction. Therefore, additional SMN-independent compounds, supporting SMN-dependent therapies, might be beneficial. Neurocalcin delta (NCALD) reduction, an SMA protective genetic modifier, ameliorates SMA across species. In a low-dose SMN-ASO-treated severe SMA mouse model, presymptomatic intracerebroventricular (i.c.v.) injection of Ncald-ASO at postnatal day 2 (PND2) significantly ameliorates histological and electrophysiological SMA hallmarks at PND21. However, contrary to SMN-ASOs, Ncald-ASOs show a shorter duration of action limiting a long-term benefit. Here, we investigated the longer-term effect of Ncald-ASOs by additional i.c.v. bolus injection at PND28. Two weeks after injection of 500 µg Ncald-ASO in wild-type mice, NCALD was significantly reduced in the brain and spinal cord and well tolerated. Next, we performed a double-blinded preclinical study combining low-dose SMN-ASO (PND1) with 2× i.c.v. Ncald-ASO or CTRL-ASO (100 µg at PND2, 500 µg at PND28). Ncald-ASO re-injection significantly ameliorated electrophysiological defects and NMJ denervation at 2 months. Moreover, we developed and identified a non-toxic and highly efficient human NCALD-ASO that significantly reduced NCALD in hiPSC-derived MNs. This improved both neuronal activity and growth cone maturation of SMA MNs, emphasizing the additional protective effect of NCALD-ASO treatment. Full article

(This article belongs to the Special Issue Regeneration for Spinal Diseases 3.0)

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23 pages, 5962 KiB

Open AccessArticle

An Injectable Engineered Cartilage Gel Improves Intervertebral Disc Repair in a Rat Nucleotomy Model

by Basanta Bhujel, Soon Shim Yang, Hwal Ran Kim, Sung Bum Kim, Byoung-Hyun Min, Byung Hyune Choi and Inbo Han

Int. J. Mol. Sci. 2023, 24(4), 3146; https://doi.org/10.3390/ijms24043146 - 5 Feb 2023

Cited by 4 | Viewed by 3626

Abstract

Lower back pain is a major problem caused by intervertebral disc degeneration. A common surgical procedure is lumbar partial discectomy (excision of the herniated disc causing nerve root compression), which results in further disc degeneration, severe lower back pain, and disability after discectomy. Thus, the development of disc regenerative therapies for patients who require lumbar partial discectomy is crucial. Here, we investigated the effectiveness of an engineered cartilage gel utilizing human fetal cartilage-derived progenitor cells (hFCPCs) on intervertebral disc repair in a rat tail nucleotomy model. Eight-week-old female Sprague-Dawley rats were randomized into three groups to undergo intradiscal injection of (1) cartilage gel, (2) hFCPCs, or (3) decellularized extracellular matrix (ECM) (n = 10/each group). The treatment materials were injected immediately after nucleotomy of the coccygeal discs. The coccygeal discs were removed six weeks after implantation for radiologic and histological analysis. Implantation of the cartilage gel promoted degenerative disc repair compared to hFCPCs or hFCPC-derived ECM by increasing the cellularity and matrix integrity, promoting reconstruction of nucleus pulposus, restoring disc hydration, and downregulating inflammatory cytokines and pain. Our results demonstrate that cartilage gel has higher therapeutic potential than its cellular or ECM component alone, and support further translation to large animal models and human subjects. Full article

(This article belongs to the Special Issue Regeneration for Spinal Diseases 3.0)

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17 pages, 3626 KiB

Open AccessArticle

Concentration-Dependent Efficacy of Recombinant Human Bone Morphogenetic Protein-2 Using a HA/β-TCP Hydrogel Carrier in a Mini-Pig Vertebral Oblique Lateral Interbody Fusion Model

by Hye-Yeong Lee, Ji-In Kang, Hye-Lan Lee, Gwang-Yong Hwang, Keung-Nyun Kim and Yoon Ha

Int. J. Mol. Sci. 2023, 24(1), 892; https://doi.org/10.3390/ijms24010892 - 3 Jan 2023

Cited by 2 | Viewed by 2783

Abstract

Bone morphogenetic protein-2 (BMP-2) is used in the treatment of degenerative spinal disease and vertebral fractures, spine fusion, dental surgery, and facial surgery. However, high doses are associated with side effects such as inflammation and osteophytes. In this study, we performed spinal fusion surgery on mini-pigs using BMP-2 and a HA/β-TCP hydrogel carrier, and evaluated the degree of fusion and osteophyte growth according to time and dosage. Increasing the dose of BMP-2 led to a significantly higher fusion rate than was observed in the control group, and there was no significant difference between the 8-week and 16-week samples. We also found that the HA + β-TCP hydrogel combination helped maintain the rate of BMP-2 release. In conclusion, the BMP-2-loaded HA/β-TCP hydrogel carrier used in this study overcame the drawback of potentially causing side effects when used at high concentrations by enabling the sustained release of BMP-2. This method is also highly efficient, since it provides mineral matter to accelerate the fusion rate of the spine and improve bone quality. Full article

(This article belongs to the Special Issue Regeneration for Spinal Diseases 3.0)

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Review

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20 pages, 635 KiB

Open AccessReview

Advances in Platelet-Rich Plasma Treatment for Spinal Diseases: A Systematic Review

by Soya Kawabata, Koji Akeda, Junichi Yamada, Norihiko Takegami, Tatsuhiko Fujiwara, Nobuyuki Fujita and Akihiro Sudo

Int. J. Mol. Sci. 2023, 24(8), 7677; https://doi.org/10.3390/ijms24087677 - 21 Apr 2023

Cited by 20 | Viewed by 5693

Abstract

Spinal diseases are commonly associated with pain and neurological symptoms, which negatively impact patients’ quality of life. Platelet-rich plasma (PRP) is an autologous source of multiple growth factors and cytokines, with the potential to promote tissue regeneration. Recently, PRP has been widely used for the treatment of musculoskeletal diseases, including spinal diseases, in clinics. Given the increasing popularity of PRP therapy, this article examines the current literature for basic research and emerging clinical applications of this therapy for treating spinal diseases. First, we review in vitro and in vivo studies, evaluating the potential of PRP in repairing intervertebral disc degeneration, promoting bone union in spinal fusion surgeries, and aiding in neurological recovery from spinal cord injury. Second, we address the clinical applications of PRP in treating degenerative spinal disease, including its analgesic effect on low back pain and radicular pain, as well as accelerating bone union during spinal fusion surgery. Basic research demonstrates the promising regenerative potential of PRP, and clinical studies have reported on the safety and efficacy of PRP therapy for treating several spinal diseases. Nevertheless, further high-quality randomized controlled trials would be required to establish clinical evidence of PRP therapy. Full article

(This article belongs to the Special Issue Regeneration for Spinal Diseases 3.0)

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