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A preliminary study of osteochondral regeneration using a scaffold-free three-dimensional construct of porcine adipose tissue-derived mesenchymal stem cells.

Murata D, Tokunaga S, Tamura T, Kawaguchi H, Miyoshi N, Fujiki M, Nakayama K, Misumi K - J Orthop Surg Res (2015)

Bottom Line: The histopathology of the implants after 6 months revealed active endochondral ossification underneath the plump fibrocartilage in animal no. 1.The histopathology after 12 months in animal no. 2 showed not only that the diminishing fibrocartilage was as thick as the surrounding normal cartilage but also that massive subchondral bone was present.The present results suggest that implantation of a scaffold-free 3D construct of AT-MSCs into an osteochondral defect may induce regeneration of the original structure of the cartilage and subchondral bone over the course of 1 year, although more experimental cases are needed.

View Article: PubMed Central - PubMed

Affiliation: Veterinary Surgery, Department of Veterinary Clinical Science, Joint Faculty of Veterinary Medicine, Kagoshima University, 21-24 Korimoto 1-chome, Kagoshima, 890-0065, Japan. daiki_net_offficial@yahoo.co.jp.

ABSTRACT

Background: Osteoarthritis (OA) is a major joint disease in humans and many other animals. Consequently, medical countermeasures for OA have been investigated diligently. This study was designed to examine the regeneration of articular cartilage and subchondral bone using three-dimensional (3D) constructs of adipose tissue-derived mesenchymal stem cells (AT-MSCs).

Methods: AT-MSCs were isolated and expanded until required for genetical and immunological analysis and construct creation. A construct consisting of about 760 spheroids that each contained 5.0 × 10(4) autologous AT-MSCs was implanted into an osteochondral defect (diameter: 4 mm; depth: 6 mm) created in the femoral trochlear groove of two adult microminipigs. After implantation, the defects were monitored by computed tomography every month for 6 months in animal no. 1 and 12 months in animal no. 2.

Results: AT-MSCs were confirmed to express the premature genes and to be positive for CD90 and CD105 and negative for CD34 and CD45. Under specific nutrient conditions, the AT-MSCs differentiated into osteogenic, chondrogenic, and adipogenic lineages, as evidenced by the expressions of related marker genes and the production of appropriate matrix molecules. A radiopaque area emerged from the boundary between the bone and the implant and increased more steadily upward and inward for the implants in both animal no. 1 and animal no. 2. The histopathology of the implants after 6 months revealed active endochondral ossification underneath the plump fibrocartilage in animal no. 1. The histopathology after 12 months in animal no. 2 showed not only that the diminishing fibrocartilage was as thick as the surrounding normal cartilage but also that massive subchondral bone was present.

Conclusions: The present results suggest that implantation of a scaffold-free 3D construct of AT-MSCs into an osteochondral defect may induce regeneration of the original structure of the cartilage and subchondral bone over the course of 1 year, although more experimental cases are needed.

No MeSH data available.


Related in: MedlinePlus

Surgical procedure. A columnar construct (4 mm in diameter and 6 mm in height) for the implantation (A). A cylindrical osteochondral defect in each groove before implantation (B). The construct composed of about 760 spheroids of AT-MSCs was autografted into the osteochondral defect in the right hind limb (C). Nothing was implanted into the left limbs (control defects; B).
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Fig1: Surgical procedure. A columnar construct (4 mm in diameter and 6 mm in height) for the implantation (A). A cylindrical osteochondral defect in each groove before implantation (B). The construct composed of about 760 spheroids of AT-MSCs was autografted into the osteochondral defect in the right hind limb (C). Nothing was implanted into the left limbs (control defects; B).

Mentions: At least 4 × 107 AT-MSCs were used to produce each autologous construct. The cells were inoculated into eight 96-well plates (Sumitomo Bakelite, Tokyo, Japan) with 5 × 104 cells/well. After undisrupted incubation for 48 h, the cells formed spheroids with a diameter of about 700 μm in the bottom of the wells. About 760 spheroids were placed in a cylindrical mold and incubated in CCM until implantation (7 days). When the mold was carefully removed, a columnar construct of 4 mm in diameter and 6 mm in height appeared and was used for autologous implantation (Figure 1A). The general outline of this method of construction has already been reported [21,33].Figure 1


A preliminary study of osteochondral regeneration using a scaffold-free three-dimensional construct of porcine adipose tissue-derived mesenchymal stem cells.

Murata D, Tokunaga S, Tamura T, Kawaguchi H, Miyoshi N, Fujiki M, Nakayama K, Misumi K - J Orthop Surg Res (2015)

Surgical procedure. A columnar construct (4 mm in diameter and 6 mm in height) for the implantation (A). A cylindrical osteochondral defect in each groove before implantation (B). The construct composed of about 760 spheroids of AT-MSCs was autografted into the osteochondral defect in the right hind limb (C). Nothing was implanted into the left limbs (control defects; B).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4389925&req=5

Fig1: Surgical procedure. A columnar construct (4 mm in diameter and 6 mm in height) for the implantation (A). A cylindrical osteochondral defect in each groove before implantation (B). The construct composed of about 760 spheroids of AT-MSCs was autografted into the osteochondral defect in the right hind limb (C). Nothing was implanted into the left limbs (control defects; B).
Mentions: At least 4 × 107 AT-MSCs were used to produce each autologous construct. The cells were inoculated into eight 96-well plates (Sumitomo Bakelite, Tokyo, Japan) with 5 × 104 cells/well. After undisrupted incubation for 48 h, the cells formed spheroids with a diameter of about 700 μm in the bottom of the wells. About 760 spheroids were placed in a cylindrical mold and incubated in CCM until implantation (7 days). When the mold was carefully removed, a columnar construct of 4 mm in diameter and 6 mm in height appeared and was used for autologous implantation (Figure 1A). The general outline of this method of construction has already been reported [21,33].Figure 1

Bottom Line: The histopathology of the implants after 6 months revealed active endochondral ossification underneath the plump fibrocartilage in animal no. 1.The histopathology after 12 months in animal no. 2 showed not only that the diminishing fibrocartilage was as thick as the surrounding normal cartilage but also that massive subchondral bone was present.The present results suggest that implantation of a scaffold-free 3D construct of AT-MSCs into an osteochondral defect may induce regeneration of the original structure of the cartilage and subchondral bone over the course of 1 year, although more experimental cases are needed.

View Article: PubMed Central - PubMed

Affiliation: Veterinary Surgery, Department of Veterinary Clinical Science, Joint Faculty of Veterinary Medicine, Kagoshima University, 21-24 Korimoto 1-chome, Kagoshima, 890-0065, Japan. daiki_net_offficial@yahoo.co.jp.

ABSTRACT

Background: Osteoarthritis (OA) is a major joint disease in humans and many other animals. Consequently, medical countermeasures for OA have been investigated diligently. This study was designed to examine the regeneration of articular cartilage and subchondral bone using three-dimensional (3D) constructs of adipose tissue-derived mesenchymal stem cells (AT-MSCs).

Methods: AT-MSCs were isolated and expanded until required for genetical and immunological analysis and construct creation. A construct consisting of about 760 spheroids that each contained 5.0 × 10(4) autologous AT-MSCs was implanted into an osteochondral defect (diameter: 4 mm; depth: 6 mm) created in the femoral trochlear groove of two adult microminipigs. After implantation, the defects were monitored by computed tomography every month for 6 months in animal no. 1 and 12 months in animal no. 2.

Results: AT-MSCs were confirmed to express the premature genes and to be positive for CD90 and CD105 and negative for CD34 and CD45. Under specific nutrient conditions, the AT-MSCs differentiated into osteogenic, chondrogenic, and adipogenic lineages, as evidenced by the expressions of related marker genes and the production of appropriate matrix molecules. A radiopaque area emerged from the boundary between the bone and the implant and increased more steadily upward and inward for the implants in both animal no. 1 and animal no. 2. The histopathology of the implants after 6 months revealed active endochondral ossification underneath the plump fibrocartilage in animal no. 1. The histopathology after 12 months in animal no. 2 showed not only that the diminishing fibrocartilage was as thick as the surrounding normal cartilage but also that massive subchondral bone was present.

Conclusions: The present results suggest that implantation of a scaffold-free 3D construct of AT-MSCs into an osteochondral defect may induce regeneration of the original structure of the cartilage and subchondral bone over the course of 1 year, although more experimental cases are needed.

No MeSH data available.


Related in: MedlinePlus