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The effect of poloxamer 407-based hydrogel on the osteoinductivity of demineralized bone matrix.

Lee JH, Baek HR, Lee KM, Lee HK, Im SB, Kim YS, Lee JH, Chang BS, Lee CK - Clin Orthop Surg (2014)

Bottom Line: Both of the compositions were applied to human mesenchymal stem cell (MSC) cultures, and monitored for alkaline phosphatase (ALP) staining and ALP activity.The DBM-W group showed stronger ALP staining at 7, 14, and 21 days of treatment, and significantly higher ALP activity at 7 and 14 days of treatment, compared to the DBM-H group.Decalcified histology found bony tissue with lamellation in both groups.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedic Surgery, SMG-SNU Boramae Medical Center, Seoul National University College of Medicine, Seoul, Korea. ; Institute of Medical and Biological Engineering, Medical Research Center, Seoul National University, Seoul, Korea.

ABSTRACT

Background: Demineralized bone matrix (DBM) is used for bone healing due to its osteoinductivity, but it requires a carrier for clinical application. Here, we report the effects on the osteoinductivity of DBM by use of a poloxamer 407-based hydrogel as the carrier, compared to sterile water.

Methods: DBM-W and DBM-H represent 27 wt% of DBM with sterile water and DBM with a poloxamer 407-based hydrogel, respectively. Both of the compositions were applied to human mesenchymal stem cell (MSC) cultures, and monitored for alkaline phosphatase (ALP) staining and ALP activity. Six 10-week-old athymic nude rats were used for abdominal muscle grafting with either DBM-W or DBM-H, and were tested by plane radiography, microfocus X-ray computed tomography (CT), and decalcified histology to evaluate ectopic bone formation.

Results: The DBM-W group showed stronger ALP staining at 7, 14, and 21 days of treatment, and significantly higher ALP activity at 7 and 14 days of treatment, compared to the DBM-H group. Plane radiography could not confirm the radio-opaque lesions in the rat ectopic bone formulation model. However, ectopic bone formation was observed in both groups by micro-CT. Compared to the DBM-H group, the DBM-W group showed higher bone volume, percent bone volume and trabecular number, and the difference in percent bone volume was statistically significant. Decalcified histology found bony tissue with lamellation in both groups.

Conclusions: Our results suggest that poloxamer 407-based hydrogel has efficacy as a DBM carrier since it shows ectopic bone formation, but its effects on the quality and quantity of osteoblastic differentiation in rat abdominal ectopic bone and MSC are considered negative.

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Related in: MedlinePlus

Operation process. Three incisions and pouches were made per side (right and left) at the abdominal muscle of athymic nude rats. (A) A picture of fixed demineralized bone matrix (DBM). (B) A picture of the sutured pouches after DBM implantation.
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Figure 1: Operation process. Three incisions and pouches were made per side (right and left) at the abdominal muscle of athymic nude rats. (A) A picture of fixed demineralized bone matrix (DBM). (B) A picture of the sutured pouches after DBM implantation.

Mentions: A total of 6 male athymic nude rats (Hsd:RH-Foxn1, 10 weeks, 280-290 g) were used for this study, with approval from the Standing Ethical Committee at the Laboratory for Animal Research in the Clinical Research Institute of the Seoul National University Hospital (IACUC No. 10-0083). This study followed the 'Guiding Principles for Research Involving Animals and Human Beings' by the American Physiological Society. Rats in this study were given at least a week for adaptation. They were housed in a specific pathogen-free environment with a 12-hour light/12-hour dark cycle at 24℃. Sterile water was available at all times as drinking water, and the animals were fed commercial diets. Three out of 6 rats were randomly selected for the test with DBM-W, and the rest were assigned to the DBM-H group. Zoletil (0.4 mL/kg, Virbac Laboratories, Carros, France) and rompun (10 mg/kg, Bayer Korea Ltd., Seoul, Korea) were used for intraperitoneal anesthesia. After depilation and skin preparation, a longitudinal skin incision was made in the center of the abdominal region. Three pouches were made in the right and left sides of the abdominal muscle. One milliliter of DBM was inserted into the pouches, and the facial layer and skin were sutured (Fig. 1). Immediately after the operation, the rats were intravenously administered with 100 mg of cephazolin, and were then raised without any intervention. Eight weeks later, the animals were sacrificed with an intravenous injection of KCl, after abdominal anesthesia with zoletil and rompun for further tests.


The effect of poloxamer 407-based hydrogel on the osteoinductivity of demineralized bone matrix.

Lee JH, Baek HR, Lee KM, Lee HK, Im SB, Kim YS, Lee JH, Chang BS, Lee CK - Clin Orthop Surg (2014)

Operation process. Three incisions and pouches were made per side (right and left) at the abdominal muscle of athymic nude rats. (A) A picture of fixed demineralized bone matrix (DBM). (B) A picture of the sutured pouches after DBM implantation.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4233226&req=5

Figure 1: Operation process. Three incisions and pouches were made per side (right and left) at the abdominal muscle of athymic nude rats. (A) A picture of fixed demineralized bone matrix (DBM). (B) A picture of the sutured pouches after DBM implantation.
Mentions: A total of 6 male athymic nude rats (Hsd:RH-Foxn1, 10 weeks, 280-290 g) were used for this study, with approval from the Standing Ethical Committee at the Laboratory for Animal Research in the Clinical Research Institute of the Seoul National University Hospital (IACUC No. 10-0083). This study followed the 'Guiding Principles for Research Involving Animals and Human Beings' by the American Physiological Society. Rats in this study were given at least a week for adaptation. They were housed in a specific pathogen-free environment with a 12-hour light/12-hour dark cycle at 24℃. Sterile water was available at all times as drinking water, and the animals were fed commercial diets. Three out of 6 rats were randomly selected for the test with DBM-W, and the rest were assigned to the DBM-H group. Zoletil (0.4 mL/kg, Virbac Laboratories, Carros, France) and rompun (10 mg/kg, Bayer Korea Ltd., Seoul, Korea) were used for intraperitoneal anesthesia. After depilation and skin preparation, a longitudinal skin incision was made in the center of the abdominal region. Three pouches were made in the right and left sides of the abdominal muscle. One milliliter of DBM was inserted into the pouches, and the facial layer and skin were sutured (Fig. 1). Immediately after the operation, the rats were intravenously administered with 100 mg of cephazolin, and were then raised without any intervention. Eight weeks later, the animals were sacrificed with an intravenous injection of KCl, after abdominal anesthesia with zoletil and rompun for further tests.

Bottom Line: Both of the compositions were applied to human mesenchymal stem cell (MSC) cultures, and monitored for alkaline phosphatase (ALP) staining and ALP activity.The DBM-W group showed stronger ALP staining at 7, 14, and 21 days of treatment, and significantly higher ALP activity at 7 and 14 days of treatment, compared to the DBM-H group.Decalcified histology found bony tissue with lamellation in both groups.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedic Surgery, SMG-SNU Boramae Medical Center, Seoul National University College of Medicine, Seoul, Korea. ; Institute of Medical and Biological Engineering, Medical Research Center, Seoul National University, Seoul, Korea.

ABSTRACT

Background: Demineralized bone matrix (DBM) is used for bone healing due to its osteoinductivity, but it requires a carrier for clinical application. Here, we report the effects on the osteoinductivity of DBM by use of a poloxamer 407-based hydrogel as the carrier, compared to sterile water.

Methods: DBM-W and DBM-H represent 27 wt% of DBM with sterile water and DBM with a poloxamer 407-based hydrogel, respectively. Both of the compositions were applied to human mesenchymal stem cell (MSC) cultures, and monitored for alkaline phosphatase (ALP) staining and ALP activity. Six 10-week-old athymic nude rats were used for abdominal muscle grafting with either DBM-W or DBM-H, and were tested by plane radiography, microfocus X-ray computed tomography (CT), and decalcified histology to evaluate ectopic bone formation.

Results: The DBM-W group showed stronger ALP staining at 7, 14, and 21 days of treatment, and significantly higher ALP activity at 7 and 14 days of treatment, compared to the DBM-H group. Plane radiography could not confirm the radio-opaque lesions in the rat ectopic bone formulation model. However, ectopic bone formation was observed in both groups by micro-CT. Compared to the DBM-H group, the DBM-W group showed higher bone volume, percent bone volume and trabecular number, and the difference in percent bone volume was statistically significant. Decalcified histology found bony tissue with lamellation in both groups.

Conclusions: Our results suggest that poloxamer 407-based hydrogel has efficacy as a DBM carrier since it shows ectopic bone formation, but its effects on the quality and quantity of osteoblastic differentiation in rat abdominal ectopic bone and MSC are considered negative.

Show MeSH
Related in: MedlinePlus