Simultaneous gene transfer of bone morphogenetic protein (BMP)-2 and BMP-7 by in vivo electroporation induces rapid bone formation and BMP-4 expression

Background: Transcutaneous in vivo electroporation is expected to be an effective gene-transfer method for promoting bone regeneration using the BMP-2 plasmid vector. To promote enhanced osteoinduction using this method, we simultaneously transferred cDNAs for BMP-2 and BMP-7, as inserts in the non-viral vector pCAGGS. Methods: First, an in vitro study was carried out to confirm the expression of BMP-2 and BMP-7 following the double-gene transfer. Next, the individual BMP-2 and BMP-7 plasmids or both together were injected into rat calf muscles, and transcutaneous electroporation was applied 8 times at 100 V, 50 msec. Results: In the culture system, the simultaneous transfer of the BMP-2 and BMP-7 genes led to a much higher ALP activity in C2C12 cells than did the transfer of either gene alone. In vivo, ten days after the treatment, soft X-ray analysis showed that muscles that received both pCAGGS-BMP-2 and pCAGGS-BMP-7 had better-defined opacities than those receiving a single gene. Histological examination showed advanced ossification in calf muscles that received the double-gene transfer. BMP-4 mRNA was also expressed, and RT-PCR showed that its level increased for 3 days in a timedependent manner in the double-gene transfer group. Immunohistochemistry confirmed that BMP- 4-expressing cells resided in the matrix between muscle fibers. Conclusion: The simultaneous transfer of BMP-2 and BMP-7 genes using in vivo electroporation induces more rapid bone formation than the transfer of either gene alone, and the increased expression of endogenous BMP-4 suggests that the rapid ossification is related to the induction of BMP-4.</p

Periodontal Tissue as a Biomaterial for Hard-Tissue Regeneration following bmp-2 Gene Transfer

The application of periodontal tissue in regenerative medicine has gained increasing interest since it has a high potential to induce hard-tissue regeneration, and is easy to handle and graft to other areas of the oral cavity or tissues. Additionally, bone morphogenetic protein-2 (BMP-2) has a high potential to induce the differentiation of mesenchymal stem cells into osteogenic cells. We previously developed a system for a gene transfer to the periodontal tissues in animal models. In this study, we aimed to reveal the potential and efficiency of periodontal tissue as a biomaterial for hard-tissue regeneration following a bmp-2 gene transfer. A non-viral expression vector carrying bmp-2 was injected into the palate of the periodontal tissues of Wistar rats, followed by electroporation. The periodontal tissues were analyzed through bone morphometric analyses, including mineral apposition rate (MAR) determination and collagen micro-arrangement, which is a bone quality parameter, before and after a gene transfer. The MAR was significantly higher 3–6 d after the gene transfer than that before the gene transfer. Collagen orientation was normally maintained even after the bmp-2 gene transfer, suggesting that the bmp-2 gene transfer has no adverse effects on bone quality. Our results suggest that periodontal tissue electroporated with bmp-2 could be a novel biomaterial candidate for hard-tissue regeneration therapy.Kawai M.Y., Ozasa R., Ishimoto T., et al. Periodontal Tissue as a Biomaterial for Hard-Tissue Regeneration following bmp-2 Gene Transfer. Materials, 15, 3, 993. https://doi.org/10.3390/ma15030993

Analysis of mineral apposition rates during alveolar bone regeneration over three weeks following transfer of BMP-2/7 gene via in vivo electroporation

Alveolar bone is not spontaneously regenerated following trauma or periodontitis. We previously proposed an animal model for new alveolar bone regeneration therapy based on the non-viral BMP-2/7 gene expression vector and in vivo electroporation, which induced the formation of new alveolar bone over the course of a week. Here, we analysed alveolar bone during a period of three weeks following gene transfer to periodontal tissue. Non-viral plasmid vector pCAGGS-BMP-2/7 or pCAGGS control was injected into palatal periodontal tissue of the first molar of the rat maxilla and immediately electroporated with 32 pulses of 50 V for 50 msec. Over the following three weeks, rats were double bone-stained by calcein and tetracycline every three days and mineral apposition rates (MAR) were measured. Double bone-staining revealed that MAR of alveolar bone was as similar level three days before BMP-2/7 gene transfer as three days after gene transfer. However, from 3 to 6 days, 6 to 9 days, 9 to 12 days, 12 to 15 days, 15 to 18 days, and 18 to 20 days after, MARs were significantly higher than prior to gene transfer. Our proposed gene therapy for alveolar bone regeneration combining non-viral BMP-2/7 gene expression vector and in vivo electroporation could increase alveolar bone regeneration potential in the targeted area for up to three weeks

Carbon ion radiotherapy for basal cell adenocarcinoma of the head and neck: preliminary report of six cases and review of the literature

<p>Abstract</p> <p>Background</p> <p>Basal cell adenocarcinoma accounts for approximately 1.6% of all salivary gland neoplasms. In this report, we describe our experiences of treatment for BCAC with carbon ion radiotherapy in our institution.</p> <p>Methods</p> <p>Case records of 6 patients with diagnosis of basal cell adenocarcinoma of the head and neck, who were treated by carbon ion radiotherapy with 64.0 GyE/16 fractions in our institution, were retrospectively reviewed.</p> <p>Results</p> <p>In a mean follow-up period of 32.1 months (14.0-51.3 months), overall survival and local control rates of 100% were achieved. Only one grade 4 (CTCAE v3.0) late complication occurred. There was no other grade 3 or higher toxicity.</p> <p>Conclusions</p> <p>Carbon ion radiotherapy should be considered as an appropriate curative approach for treatment of basal cell adenocarcinoma in certain cases, particularly in cases of unresectable disease and postoperative gross residual or recurrent disease.</p

Head and Neck Tumors

Clinical trial with carbon ion radiotherapy for the head and neck tumors was conducted under [the Phase I/II Clinical Trial (Protocol 9301) on Heavy Particle Radiotherapy for Malignant Head and Neck Tumors], that was initiated in June 1994 by way of a dose escalation study on a fractionation method of 18 fractions over 6 weeks. This trial was followed by a next dose fractionation and dose escalation study commenced in June 1996 under [the Phase I/II Clinical Trial (Protocol 9504) on Heavy Particle Radiotherapy for Malignant Head and Neck Tumors] using a fractionation method of 16 fractions over 4 weeks. The results of these two trials were published in 1994. Following the outcome of these two studies, the [Phase II Clinical Trial on Heavy Particle Radiotherapy for Malignant Head and Neck Tumors (Protocol 9602)] was initiated on a 64.0GyE/16 fractions/4 weeks fractionation method (or 57.6 GyE/16 fractions/4 weeks when the wide-range of the skin was included in the target volume) in April 1997.Based on the results of preliminary analysis of the 9602 protocol, two protocol were derived with effect from April 2001 into 1) the [Phase I/II Clinical Trial of Carbon Ion Radiotherapy for Bone and Soft Tissue Sarocomas in Head and Neck (Protocol 0006)] designed as a dose escalation study for bone and soft-tissue tumors, and 2) the [Phase II Clinical Trial of Carbon Ion Radiotherapy Combined with Chemotherapy for Mucosal Malignant Melanoma in Head and Neck (Protocol 0007)] for the treatment of malignant melanoma with concomitant chemotherapy.NIRS-IMP Joint Symposium on Carbon Ion Therap

Head and Neck Tumors

Clinical trial with carbon ion radiotherapy for the head and neck tumors was conducted under the Phase I/II Clinical Trial (Protocol 9301) on Heavy Particle Radiotherapy for Malignant Head and Neck Tumors, that was initiated in June 1994 by way of a dose escalation study on a fractionation method of 18 fractions over 6 weeks. This trial was followed by a next dose fractionation and dose escalation study commenced in June 1996 under the Phase I/II Clinical Trial (Protocol 9504) on Heavy Particle Radiotherapy for Malignant Head and Neck Tumors using a fractionation method of 16 fractions over 4 weeks. The results of these two trials were published in 1994. Following the outcome of these two studies, the Phase II Clinical Trial on Heavy Particle Radiotherapy for Malignant Head and Neck Tumors (Protocol 9602) was initiated on a 64.0GyE/16 fractions/4 weeks fractionation method (or 57.6 GyE/16 fractions/4 weeks when the wide-range of the skin was included in the target volume) in April 1997.Based on the results of preliminary analysis of the 9602 protocol, two protocol were derived with effect from April 2001 into 1) the Phase I/II Clinical Trial of Carbon Ion Radiotherapy for Bone and Soft Tissue Sarocomas in Head and Neck (Protocol 0006) designed as a dose escalation study for bone and soft-tissue tumors, and 2) the Phase II Clinical Trial of Carbon Ion Radiotherapy Combined with Chemotherapy for Mucosal Malignant Melanoma in Head and Neck (Protocol 0007) for the treatment of malignant melanoma with concomitant chemotherapy

Head and Neck Tumors

Clinical trial with carbon ion radiotherapy for the head and neck tumors was conducted under the Phase I/II Clinical Trial (Protocol 9301) on Heavy Particle Radiotherapy for Malignant Head and Neck Tumors, that was initiated in June 1994 by way of a dose escalation study on a fractionation method of 18 fractions over 6 weeks. This trial was followed by a next dose fractionation and dose escalation study commenced in June 1996 under the Phase I/II Clinical Trial (Protocol 9504) on Heavy Particle Radiotherapy for Malignant Head and Neck Tumors using a fractionation method of 16 fractions over 4 weeks. The results of these two trials were published in 1994 [1]. Following the outcome of these two studies, the Phase II Clinical Trial on Heavy Particle Radiotherapy for Malignant Head and Neck Tumors (Protocol 9602) was initiated on a 64.0GyE/16 fractions/4 weeks fractionation method (or 57.6 GyE/16 fractions/4 weeks when the wide-range of the skin was included in the target volume) in April 1997.Based on the results of preliminary analysis of the 9602 protocol, two protocol were derived with effect from April 2001 into 1) the Phase I/II Clinical Trial of Carbon Ion Radiotherapy for Bone and Soft Tissue Tumors of the Head and Neck (Protocol 0006) designed as a dose escalation study for bone and soft-tissue tumors, and 2) the Phase II Clinical Trial of Carbon Ion Radiotherapy Combined with Chemotherapy for Mucosal Malignant Melanoma of the Head and Neck (Protocol 0007) for the treatment of malignant melanoma with concomitant chemotherapy. The 9602 and 0007 protocols have been carried out since November 2003 under the Highly Advanced Medical Technology

Head and Neck Tumor

Clinical trial with carbon ion radiotherapy for the head and neck tumors was conducted under the Phase I/II Clinical Trial (Protocol 9301) on Heavy Particle Radiotherapy for Malignant Head and Neck Tumors, that was initiated in June 1994 by way of a dose escalation study on a fractionation method of 18 fractions over 6 weeks. This trial was followed by a next dose fractionation and dose escalation study commenced in June 1996 under the Phase I/II Clinical Trial (Protocol 9504) on Heavy Particle Radiotherapy for Malignant Head and Neck Tumors using a fractionation method of 16 fractions over 4 weeks. The results of these two trials were published in 1994 [1]. Following the outcome of these two studies, the Phase II Clinical Trial on Heavy Particle Radiotherapy for Malignant Head and Neck Tumors (Protocol 9602) was initiated on a 64.0GyE/16 fractions/4 weeks fractionation method (or 57.6 GyE/16 fractions/4 weeks when the wide-range of the skin was included in the target volume) in April 1997.Based on the results of preliminary analysis of the 9602 protocol, two protocol were derived with effect from April 2001 into 1) the Phase I/II Clinical Trial of Carbon Ion Radiotherapy for Bone and Soft Tissue Tumors of the Head and Neck (Protocol 0006) designed as a dose escalation study for bone and soft-tissue tumors, and 2) the Phase II Clinical Trial of Carbon Ion Radiotherapy Combined with Chemotherapy for Mucosal Malignant Melanoma of the Head and Neck (Protocol 0007) for the treatment of malignant melanoma with concomitant chemotherapy. The 9602 and 0007 protocols have been carried out since November 2003 under the Highly Advanced Medical Technology.NIRS-ETOILE Joint Symposium　2009 on Carbon Ion Radiotherap