Mesenchymal Stem Cell-Seeded High-Density Collagen Gel for Annular Repair: 6-Week Results From In Vivo Sheep Models

Neurosurgery ◽  
2018 ◽  
Vol 85 (2) ◽  
pp. E350-E359 ◽  
Author(s):  
Ibrahim Hussain ◽  
Stephen R Sloan ◽  
Christoph Wipplinger ◽  
Rodrigo Navarro-Ramirez ◽  
Micaella Zubkov ◽  
...  
Keyword(s):  
Nucleus Pulposus ◽  
Annulus Fibrosus ◽  
Perioperative Complications ◽  
Collagen Gel ◽  
Intervertebral Discs ◽  
High Density ◽  
Sheep Model ◽  
Gel Treatment ◽  
Pfirrmann Grade

AbstractBACKGROUNDOur group has previously demonstrated in vivo annulus fibrosus repair in animal models using an acellular, riboflavin crosslinked, high-density collagen (HDC) gel.OBJECTIVETo assess if seeding allogenic mesenchymal stem cells (MSCs) into this gel yields improved histological and radiographic benefits in an in vivo sheep model of annular injury.METHODSFifteen lumbar intervertebral discs (IVDs) were randomized into 4 groups: intact, injury only, injury + acellular gel treatment, or injury + MSC-seeded gel treatment. Sheep were sacrificed at 6 wk. Disc height index (DHI), Pfirrmann grade, nucleus pulposus area, and T2 relaxation time (T2-RT) were calculated for each IVD and standardized to healthy controls from the same sheep. Quantitative histological assessment was also performed using the Han scoring system.RESULTSAll treated IVDs retained gel plugs on gross assessment and there were no adverse perioperative complications. The MSC-seeded gel treatment group demonstrated statistically significant improvement over other experimental groups in DHI (P = .002), Pfirrmann grade (P < .001), and T2-RT (P = .015). There was a trend for greater Han scores in the MSC-seeded gel-treated discs compared with injury only and acellular gel-treated IVDs (P = .246).CONCLUSIONMSC-seeded HDC gel can be delivered into injured IVDs and maintained safely in live sheep to 6 wk. Compared with no treatment and acellular HDC gel, our data show that MSC-seeded HDC gel improves outcomes in DHI, Pfirrmann grade, and T2-RT. Histological analysis shows improved annulus fibrosus and nucleus pulposus reconstitution and organization over other experimental groups as well.

scholarly journals Combined Nucleus Pulposus Augmentation and Annulus Fibrosus Repair Prevents Intervertebral Disc Degeneration After Discectomy

Neurosurgery ◽  
2019 ◽  
Vol 66 (Supplement_1) ◽  
Author(s):  
Sertac Kirnaz ◽  
Stephen Sloan ◽  
Christoph Wipplinger ◽  
Franziska Anna Schmidt ◽  
Roger Hartl ◽  
...  
Keyword(s):  
Nucleus Pulposus ◽  
Disc Degeneration ◽  
Annulus Fibrosus ◽  
Combined Treatment ◽  
Disc Height ◽  
Histological Evaluation ◽  
Sheep Model ◽  
Mri Scans ◽  
The Individual

Abstract INTRODUCTION The objective of the current study is to assess the efficacy of combined annulus fibrosus (AF) using a high-density collagen (HDC) gel and nucleus pulposus (NP) repair using a hyaluronic acid (HA) gel in an in Vivo sheep model. METHODS We performed an anterolateral, retroperitoneal prepsoas approach to access the IVDs L1-6 in a total of 8 skeletally mature Finn sheep. IVDs were randomized into 5 groups: (1) intact, (2) injured via 3 × 10 mm box annulotomy and removal of 200 mg of NP, (3) injury and HDC gel patch for AF repair, (4) injury and injection of a HA gel into the NP, and (5) injury and HDC AF repair and NP HA replacement. At 6 wk postoperatively, sheep were sacrificed and underwent postmortem 3T-MRI scans as well as gross anatomical and histological evaluation. Disc height index (DHI) analysis and Pfirrmann grading (PG) were performed on each segment using MR images. RESULTS Intact control discs were not degenerated and had an average PG of 1 while injured, and untreated discs had a significant degeneration with an average PG of 3. Discs receiving the combined injection and collagen AF patch individually showed fewer signs of degeneration than injured alone, and the combined treatment resulted in the least amount of degeneration with PG not significantly different from the intact controls. DHI confirmed the trends seen in the PG, where injured discs lost 20% of the intact disc height, the individual NP and AF repairs restored 5% to 10% of intact disc height, and the combined repairs preserved 90% of the intact disc height. CONCLUSION PG and DHI results demonstrate that individual NP and AF repairs are able to prevent disc degeneration better than no treatment at all; however, the greatest preservation of disc health was seen with combined AF and NP repairs.

Riboflavin crosslinked high-density collagen gel for the repair of annular defects in intervertebral discs: An in vivo study

2015 ◽  
Vol 26 ◽  
pp. 215-224 ◽  
Author(s):  
Peter Grunert ◽  
Brandon H. Borde ◽  
Sara B. Towne ◽  
Yu Moriguchi ◽  
Katherine D. Hudson ◽  
...  
Keyword(s):  
Collagen Gel ◽  
Intervertebral Discs ◽  
High Density ◽  
In Vivo Study

In vivo annular repair using high-density collagen gel seeded with annulus fibrosus cells

2018 ◽  
Vol 79 ◽  
pp. 230-238 ◽  
Author(s):  
Yu Moriguchi ◽  
Brandon Borde ◽  
Connor Berlin ◽  
Christoph Wipplinger ◽  
Stephen R. Sloan ◽  
...  
Keyword(s):  
Annulus Fibrosus ◽  
Collagen Gel ◽  
High Density ◽  
Annular Repair

scholarly journals Evaluation of apoptotic cell death in normal and chondrodystrophic canine intervertebral discs

2012 ◽  
Vol 2 (1) ◽  
pp. 6 ◽  
Author(s):  
Marie Klauser ◽  
Franck Forterre ◽  
Marcus Doherr ◽  
Andreas Zurbriggen ◽  
David Spreng ◽  
...  
Keyword(s):  
Nucleus Pulposus ◽  
Disc Degeneration ◽  
Annulus Fibrosus ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Intervertebral Discs ◽  
Discogenic Pain ◽  
Age Related ◽  
Chondroid Metaplasia ◽  
Related Increase

Disc degeneration occurs commonly in dogs. A variety of factors is thought to contribute an inappropriate disc matrix that isolate cells in the disc and lead to apoptosis. Disc herniation with radiculopathy and discogenic pain are the results of the degenerative process. The objective of this prospective study was to determine the extent of apoptosis in intact and herniated intervertebral discs of chondrodystrophic dogs and non-chondrodystrophic dogs. In addition, the nucleus pulposus (NP) was histologically compared between non-chondrodystrophic and chondrodystrophic dogs. Thoracolumbar intervertebral discs and parts of the extruded nucleus pulposus were harvested from 45 dogs. Samples were subsequently stained with haematoxylin-eosin and processed to detect cleaved caspase-3 and poly(ADP-ribose) polymerase. A significant greater degree of apoptosis was observed in herniated NPs of chondrodystrophic dogs compared to non- chondrodystrophic dogs with poly (ADP-ribose) polymerase and cleaved caspase- 3 detection. Within the group of chondrodystrophic dogs, dogs with an intact disc and younger than 6 years showed a significant lower incidence of apoptosis in the NP compared to the herniated NP of chondrodystrophic dogs. The extent of apoptosis in the annulus fibrosus was not different between the intact disc from chondrodystrophic and non- chondrodystrophic dogs. An age-related increase of apoptotic cells in NP and annulus fibrosus was found in the intact non-herniated intervertebral discs. Histologically, absence of notochordal cells and occurrence of chondroid metaplasia were observed in the nucleus pulposus of chondrodystrophic dogs. As a result, we found that apoptosis plays a role in disc degeneration in chondrodystrophic dogs.

scholarly journals Early differentation of lamellar structure of the intervertebral disc in staged human embryos

2015 ◽  
Vol 84 (3) ◽  
pp. 157-166
Author(s):  
Witold Woźniak ◽  
Małgorzata Grzymisławska ◽  
Joanna Łupicka ◽  
Małgorzata Bruska ◽  
Adam Piotrowski ◽  
...  
Keyword(s):  
Intervertebral Disc ◽  
Nucleus Pulposus ◽  
Annulus Fibrosus ◽  
Developmental Stages ◽  
Intervertebral Discs ◽  
Human Embryos ◽  
Lateral Zone ◽  
Vast Literature ◽  
Dense Zone

Introduction. In the vast literature concerning the development of the intervertebral discs controversies exist as to the period of differentiation and structure of the nucleus pulposus and annulus fibrosus. These controversies result from different determination of age of the investigated embryos. Aim. Using embryos from departmental collection age of which was established according to international Carnegie staging and expressed in postfertilizational days, the differentiation of the intervertebral discs was traced. Material and methods. Study was performed on 34 embryos at developmental stages 13–23 (32–56 days). Embryos were serially sectioned in sagittal, frontal and horizontal planes. Sections were stained with various histological methods and impregnated with silver.Results. Division of sclerotomes into loose cranial and dense caudal zones (sclerotomites) was observed in embryos aged 32 days (stage 13). The intervertebral disc developed from the dense zone of sclerotome and was well recognized in embryos aged 33 days (stage 14). At the end of fifth week (embryos at stage 15, 36 days) the annulus fibrosus and the nucleus pulposus were seen. The annulus fibrosus differentiated into lateral and medial zones. Within the lateral zone cells were arranged into circular rows. These rows were considered as the first stage of laminar structure. In further developmental stages the laminae occupied both zones of the annulus fibrosus.Conclusions. The intervertebral discs develop from the dense zone of the sclerotome which is evident in embryos at stage 13 (32 days). Discs differentiate in embryos aged 33 days, when the nucleus pulposus and annulus fibrosus are recognized. In embryos aged 36 days in the annulus fibrosus circular rows forming laminar arrangement are seen.

Compartmentalization of the matrix formed by nucleus pulposus and annulus fibrosus cells in alginate gel

2002 ◽  
Vol 30 (6) ◽  
pp. 874-878 ◽  
Author(s):  
E. Thonar ◽  
H. An ◽  
K. Masuda
Keyword(s):  
Nucleus Pulposus ◽  
Annulus Fibrosus ◽  
Articular Chondrocyte ◽  
Alginate Gel ◽  
Age Related ◽  
The Matrix ◽  
Disc Cells ◽  
A Cell ◽  
Associated Matrix

Intervertebral disc cells cultured in alginate gel are capable of reforming in alginate, a matrix that consists of two compartments: a rim of metabolically active cell-associated matrix and a more abundant, but metabolically less active, further removed matrix. At any one age and in most species, the cell-associated matrix formed by a nucleus pulposus or annulus fibrosus cell cultured in this way is less abundant than that formed by an articular chondrocyte. In both the cell-associated matrix and further removed matrix, the ratio of aggrecan to collagen is significantly higher in the case of nucleus pulposus than of annulus fibrosus, a feature that also distinguishes the matrices of the nucleus pulposus and annulus fibrosus in vivo. Nucleus pulposus and annulus fibrosus cells from older donors show a decreased ability to reform a cell-associated matrix rich in aggrecan. There is, however, some evidence that gene therapy and/or exposure of the cells to defined stimulatory factors can help overcome some of these age-related limitations. This contention is supported by recent evidence that nucleus pulposus and annulus fibrosus cells from adult donors can be manipulated to form, using the recently developed alginate-recovered chondrocyte system, a resilient tissue that bears many of the characteristics of the tissue in which these cells reside in vivo.

Effects of Swelling Conditions on the Compressive Properties of Nucleus Pulposus From Bovine Intervertebral Discs

2004 ◽  
Author(s):  
David T. Korda ◽  
Delphine Perie ◽  
James C. Iatridis
Keyword(s):  
Intervertebral Disc ◽  
Water Content ◽  
Nucleus Pulposus ◽  
Annulus Fibrosus ◽  
Compressive Loading ◽  
Collagen Matrix ◽  
High Water ◽  
Intervertebral Discs ◽  
High Water Content ◽  
Outer Annulus Fibrosus

The intervertebral disc provides flexibility and load support for the spine. It consists of two main regions; the outer annulus fibrosus which is a highly organized collagen matrix and the inner nucleus pulposus which (in a healthy disc) is a proteoglycan rich gelatinous material. The predominant mode of loading on the intervertebral disc is axial compression, which generates hydrostatic pressures within the disc. The high water content of the nucleus plays a major role in supporting these loads. With age and degeneration, the water content of the nucleus changes, and is believed to significantly impact its ability to bear load. The purpose of this study therefore, was to define the effects of swelling conditions (which affect disc hydration) on the material properties of the disc under compressive loading.

scholarly journals Annulus Fibrosus Repair Using High-Density Collagen Gel

Spine ◽  
2018 ◽  
Vol 43 (4) ◽  
pp. E208-E215 ◽  
Author(s):  
Brenton Pennicooke ◽  
Ibrahim Hussain ◽  
Connor Berlin ◽  
Stephen R. Sloan ◽  
Brandon Borde ◽  
...  
Keyword(s):  
Annulus Fibrosus ◽  
Collagen Gel ◽  
High Density

scholarly journals Biochemical and Immuno-Histochemical Localization of Type IIA Procollagen in Annulus Fibrosus of Mature Bovine Intervertebral Disc

2021 ◽  
Author(s):  
Audrey McAlinden ◽  
David M Hudson ◽  
Aysel A Fernandes ◽  
Soumya Ravindran ◽  
Russell J Fernandes
Keyword(s):  
Intervertebral Disc ◽  
Nucleus Pulposus ◽  
Annulus Fibrosus ◽  
Small Diameter ◽  
Type Ii Collagen ◽  
Intervertebral Discs ◽  
Matrix Composition ◽  
Type I ◽  
Type Ii ◽  
Post Translational Modification

For next generation tissue-engineered constructs and regenerative medicine to succeed clinically, the basic biology and extracellular matrix composition of tissues that these repair techniques seek to restore have to be fully determined. Using the latest reagents coupled with tried and tested methodologies, we continue to uncover previously undetected structural proteins in mature intervertebral disc. In this study we show that the ″embryonic″ type IIA procollagen isoform (containing a cysteine-rich amino propeptide) was biochemically detectable in the annulus fibrosus of both calf and mature steer intervertebral discs, but not in the nucleus pulposus where the type IIB isoform was predominantly localized. Specifically, the triple-helical type IIA procollagen isoform immunolocalized in the outer margins of the inner annulus fibrosus. Triple helical processed type II collagen exclusively localized within the inter- lamellae regions and with type IIA procollagen in the intra-lamellae regions. Mass spectrometry of the a1(II) collagen chains from the region where type IIA procollagen localized showed high 3-hydroxylation of Proline-944, a post- translational modification that is correlated with thin collagen fibrils as in the nucleus pulposus. The findings implicate small diameter fibrils of type IIA procollagen in select regions of the annulus fibrosus where it likely contributes to the organization of collagen bundles and structural properties within the type I- type II collagen transition zone.

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