Does salivary pH affect postoperative sore throat in the head-down position of general anesthesia?

BACKGROUND: Tracheal intubation sometimes causes postoperative sore throat (POST) due to laryngeal damage. However, clinical observations suggest that the environment of the oral cavity may also affect POST. OBJECTIVE: The purpose of this study was to investigate whether salivary pH in the oral cavity affects POST. METHODS: After obtaining ethical approval, informed consent was obtained from all patients. Patients who underwent surgery in the supine position were enrolled as the control group. Patients who underwent laparoscopic surgery in the head-down position were enrolled as the intervention group. Immediately before both groups of patients were anaesthetised, expelled saliva was collected, and salivary pH was measured. Immediately postoperatively, the same measurement was carried out before the patient regained consciousness. The primary outcome was the change in salivary pH. The secondary outcome was POST. In our study, POST was defined as pharyngeal and swallowing pain in the glossopharyngeal and superior laryngeal nerves. The normal distribution of pH was tested using the Shapiro-Wilk test followed by analysis using repeated-measurements and one-way analysis of variance. Statistical significance was set at p < .05. RESULTS: A total of 62 patients were enrolled, of whom two were excluded based on the exclusion criteria. Salivary pH in the intervention group was significantly lower than that in the control group. Five patients had POST in the intervention group, whereas none had POST in the control group had POST. CONCLUSION: Acidotic-shifted saliva is considered one of the causes of POST.

Does TMJ Function and Imaging Tools Help Differentiate Between Condylar Resorption and Mandibular Hypoplasia?

PURPOSE: Differentiating between bilateral condylar resorption (CR) and mandibular hypoplasia (MH) can be challenging owing to the difficulty in chronological observation and establishing standardized measurements. The purpose of the present study was to assess whether temporomandibular joint (TMJ) function can distinguish between CR and MH and clarify the essential diagnostic imaging tools for CR. MATERIALS AND METHODS: We performed a cross-sectional study of patients with mandibular retrognathia. The primary predictor variables were a clinical dysfunction score for the TMJ, mandibular plane angle (MPA), SNA angle, SNB angle, and cortical erosion score in the condylar heads. The demographic variables were age, anterior disc displacement, and previous orthodontic treatment. The anatomic variable was the condylar height (CH). The primary outcome variable was the disease status (CR or MH). The patients were divided into the CR group and MH group. The patients with CR were selected on the basis of a CH value of less than 22 mm. TMJ function was assessed using the Helkimo clinical dysfunction index. The CH on panoramic radiographs was measured using the Kjellberg method. The MPA, SNA angle, and SNB angle were analyzed using cephalometric analysis. Cortical erosion in the condylar head was assessed using computed tomography and magnetic resonance imaging. RESULTS: A total of 23 female participants were enrolled in the present study. The average clinical dysfunction score for the TMJ was 4.4 in the CR group and 0.4 in the MH group (P < .05). The average MPA was 41.2° in the CR group and 35.5° in the MH group (P < .05). CONCLUSIONS: The present investigation has shown that assessing TMJ function and analyzing MPA using a cephalometric radiograph can differentiate CR from MH.

Indications of Potassium Titanyl Phosphate Laser Therapy for Slow-Flow Vascular Malformations in Oral Region.

BACKGROUND: Indications for laser therapy for slow-flow vascular malformations in the oral and maxillofacial regions have not been clearly documented. The authors aimed to estimate the frequency of resolution of slow-flow vascular malformations and to identify risk and prognostic factors associated with resolution in potassium titanyl phosphate (KTP) laser treatment. METHODS: This study was designed as a prospective cohort study. Patients who had diagnosed slow-flow vascular malformations were continuously assigned to receive KTP laser therapy. All patients had intralesional laser photocoagulation performed under local anesthesia. Administered power of the KTP laser was fixed at 2 watts throughout the procedure in all patients. The primary endpoint was to understand the frequency of resolution of slow-flow vascular malformations in KTP laser treatment. Secondary endpoints were: treatment outcomes based on lesion size; treatment outcomes based on location; treatment outcomes based on total energy in joules; types of complications. Treatment outcomes were judged by a clinical assessment as well as reduction in lesion size on magnetic resonance imaging. RESULTS: Data were obtained from 26 patients (9 men, 17 women) with 38 lesions. The average lesion size was 13.5 ±â€Š7.7 mm. Treatment outcomes based on lesion size showed that cure and regression were obtained in lesions less than 30 mm in size. However, lesions larger than 30 mm showed no response. Lesions in the tongue and lips showed higher cure rates than in other areas. Treatment outcomes based on administered total energy in joules showed that 68% of lesions were treated and responded well at less than 400 joules. Complication rate was relatively high in the buccal mucosal lesions. Immediate postoperative complications such as necrosis were more common in high-energy administration than in low-energy administration. CONCLUSION: Our results indicated that KTP laser therapy was effective for slow-flow vascular malformations less than 30 mm in size without significant side effects.

Intravenous Dexamethasone Administration Before Orthognathic Surgery Reduces the Postoperative Edema of the Masseter Muscle: A Randomized Controlled Trial.

PURPOSE: Dexamethasone seems to suppress postoperative swelling. However, the standard administration dose of dexamethasone for bilateral sagittal split osteotomy (BSSO) has not been reported. This study focused on clarifying the most effective dose of dexamethasone for BSSO. MATERIALS AND METHODS: This research was planned as a prospective, randomized controlled, double-blind study. Patients undergoing BSSO were randomly assigned to receive intravenous preoperative dexamethasone under 3 different dose conditions: 16 mg, 8 mg, and 0 mg (control). The endpoints of this study were 1) postoperative changes in masseter muscle thickness and buccal soft tissue thickness; 2) postoperative changes in maximum incisal opening; 3) postoperative changes in sensation of the chin and lower lip region; 4) postoperative changes in blood examination findings (white blood cell count, neutrophil count, C-reactive protein level, and lymphocyte count); and 5) types of complications. Data were recorded at 2 to 4 time intervals: before surgery, postoperative day 1, postoperative day 2, and postoperative day 3. Average age, gender, average body mass index, average surgery time, and average blood loss also were examined. Data were analyzed by 1-way analysis of variance (Bonferroni multiple-comparisons test) after the Bartlett test. RESULTS: We enrolled 24 patients, including 5 men and 19 women, in this study. The rate of increase in the thickness of the masseter muscle 24 hours after BSSO was 38.4% in the 16-mg group (n = 8), 57.7% in the 8-mg group (n = 8), and 56.1% in the 0-mg group (n = 8). The rate of increase in masseter muscle thickness in the 16-mg group was significantly lower than that in the 0-mg group (P < .05). Regarding the number of lymphocytes after surgery, the 16-mg and 8-mg groups maintained preoperative levels whereas there was a reduced number of lymphocytes in the control group. No statistically significant results were obtained for the following study endpoints: postoperative changes in maximum incisal opening and postoperative changes in sensation of the chin and lower lip region. CONCLUSIONS: This investigation showed that the most effective dose of dexamethasone for BSSO is 16 mg.

Evaluation of OK-432 Injection Therapy as Possible Primary Treatment of Intraoral Ranula.

PURPOSE: A ranula is a pseudocyst caused by mucous extravasation from the sublingual gland. Recently, a sclerosing agent, OK-432 (picibanil), has been reported to be highly effective for treating lymphangioma and cervical cystic lesions. The present study assessed the effectiveness of OK-432 injection therapy for intraoral ranula to clarify whether it can be used as the primary treatment. PATIENTS AND METHODS: The present study was a retrospective clinical study of patients with intraoral ranula who received OK-432 injection therapy from 2005 to 2015. The ranula size was measured on computed tomography or magnetic resonance imaging studies. We dissolved 1 Klinische Einheit (KE) unit of OK-432 powder in normal saline equal to the aspiration volume. The primary endpoint was the treatment results. The secondary endpoints were the relation between the treatment results and the lesion length and aspiration volume. RESULTS: A total of 23 patients received OK-432 injection therapy for an intraoral ranula. The mean lesion size was 19.96 mm. The mean aspiration volume was 2.14 mL. The number of injections was 1 to 4 (mean 1.70). The treatment results were complete regression (CR) in 18 (78.2%), partial regression (PR) in 3 (13.0%), and no response (NR) in 2 (8%) patients after the last injection. The overall efficacy rate was 91.2% (21 of 23). No serious complications were observed. The lesion length and aspiration volume of the CR group was 17.38 mm and 1.40 mL, respectively. The lesion length and aspiration volume of the PR/NR group was 29.20 mm and 4.80 mL, respectively. The PR/NR group lesions were significantly larger than the CR group lesions. CONCLUSIONS: OK-432 injection therapy for intraoral ranula is safe and effective compared with other surgical therapies. This therapy could potentially become a primary treatment of intraoral ranula.

Adenosquamous carcinoma of the tongue: clinicopathologic study and review of the literature.

Adenosquamous carcinoma (ASC) of the tongue is an uncommon malignant oral neoplasm with mixed glandular and squamous differentiation and a propensity for aggressive clinical behavior. Here, we report a rare case of ASC of the lateral border of the tongue in a 65-year-old Japanese man. The patient was treated by radical operation and remained well for 6 months before developing metastasis of the hilar and pretracheal lymph nodes. Subsequently, the patient was treated with combined chemotherapy (nedaplatin plus docetaxel and S-1 for two cycles, intravenously) and radiotherapy. Radiation therapy of metastatic lymph nodes was performed at a total dose of 60 Gy and was delivered in 2 Gy fractions 5 days/week. The patient is currently tumor free and is being followed up carefully. This article describes a rare case of ASC of the tongue and its conventional histologic, immunohistochemical, and electron microscopic findings, together with a review of the literature. The findings provide important information to better understand the possible clinical and therapeutic approaches for this uncommon tumor of the tongue.

Nerve Sharing Between the Lingual and Mental Nerve to Restore Lower Lip Sensation After Segmental Resection of the Mandible.

This report demonstrates a successful new procedure for reconstructing the inferior alveolar nerve by transplanting the great auricular nerve (GAN) between the mental nerve and the remaining submandibular ganglion to achieve nerve sharing of the lingual nerve. A 59-year-old woman with discomfort in the left mandibular retromolar region and ipsilateral neck was referred to our hospital by a local dentist. Physical examination showed mild swelling and redness at the left mandibular retromolar region. The histologic diagnosis showed central mucoepidermoid carcinoma of the jaw. With the patient under general anesthesia, segmental resection of the mandible followed by level 1 selective neck dissection was performed. The resected mandible was reconstructed with a titanium plate. The submandibular incision was extended to the lower edge of the tragus for harvesting of the GAN. The GAN was grafted, and an epineural neurorrhaphy was carried out with the mental nerve, as well as the submandibular ganglion, under a microscope. After the operation, submental sensation was evaluated with a Semmes-Weinstein pressure esthesiometer. The Semmes-Weinstein pressure esthesiometer test showed a loss of perception at the third week after surgery. Within 12 months, nerve sensation was substantially improved and the patient was free from discomfort.

A randomized trial to identify the most effective dose of remifentanil during Le Fort I osteotomy.

PURPOSE: The Le Fort I osteotomy (L-I) requires extensive dissection and manipulation of tissue, causing hemodynamic instability and an undesirable postoperative stress response. This study aimed to clarify the most effective dose of remifentanil during L-I. MATERIALS AND METHODS: This study was designed as a prospective, randomized, controlled double-blinded study. Patients (American Society of Anesthesiologists physical status I to II) undergoing L-I were randomly assigned to receive anesthesia with propofol and remifentanil under 3 remifentanil dose conditions: 0.25 µg/kg/minute (group 1), 0.5 µg/kg/minute (group 2), and 0.75 µg/kg/minute (group 3). All patients underwent L-I with propofol and remifentanil target-controlled anesthesia. The study endpoints were mean arterial pressure (MAP) and heart rate. Data were recorded before L-I (20-minute period before surgery), during L-I (from the beginning of surgery to downfracture), and after L-I (20-minute period after downfracture). Average age, gender, average body mass index, aimed maxillary position, average bispectral index, average surgery time, and average blood loss also were examined. Data were analyzed using the Bartlett test and then 1-way analysis of variance with the Bonferroni multiple comparison test. RESULTS: Data were obtained from 20 patients (9 men, 11 women). The average operating times for groups 1, 2, and 3 were 53.1, 46.7, and 49 minutes, respectively. The age range was 18 to 46 years (average, 26.05 yr). The rate of MAP increase from before to during L-I in group 1 was 10.8% (n = 7). The rate of MAP increase from before to during L-I in group 3 was 2.1% (n = 6). Group 3 showed a significantly lower rate of MAP increase during and after L-I compared with group 1 (P < .05). CONCLUSION: Remifentanil administration at 0.75 µg/kg/minute stabilized hemodynamics during L-I without major side effects. Results indicated that the standard index of remifentanil administration during L-I should be 0.75 µg/kg/minute when using oxygen, propofol, and remifentanil for general anesthesia.

Tissue-engineered bone with 3-dimensionally printed ß-tricalcium phosphate and polycaprolactone scaffolds and early implantation: an in vivo pilot study in a porcine mandible model.

PURPOSE: Deep bone penetration into implanted scaffolds remains a challenge in tissue engineering. The purpose of this study was to evaluate bone penetration depth within 3-dimensionally (3D) printed ß-tricalcium phosphate (ß-TCP) and polycaprolactone (PCL) scaffolds, seeded with porcine bone marrow progenitor cells (pBMPCs), and implanted early in vivo. MATERIALS AND METHODS: Scaffolds were 3D printed with 50% ß-TCP and 50% PCL. The pBMPCs were harvested, isolated, expanded, and differentiated into osteoblasts. Cells were seeded into the scaffolds and constructs were incubated in a rotational oxygen-permeable bioreactor system for 14 days. Six 2- × 2-cm defects were created in each mandible (N = 2 minipigs). In total, 6 constructs were placed within defects and 6 defects were used as controls (unseeded scaffolds, n = 3; empty defects, n = 3). Eight weeks after surgery, specimens were harvested and analyzed by hematoxylin and eosin (H&E), 4',6-diamidino-2-phenylindole (DAPI), and CD31 staining. Analysis included cell counts, bone penetration, and angiogenesis at the center of the specimens. RESULTS: All specimens (N = 12) showed bone formation similar to native bone at the periphery. Of 6 constructs, 4 exhibited bone formation in the center. Histomorphometric analysis of the H&E-stained sections showed an average of 22.1% of bone in the center of the constructs group compared with 1.87% in the unseeded scaffolds (P < .05). The 2 remaining constructs, which did not display areas of mature bone in the center, showed massive cell penetration depth by DAPI staining, with an average of 2,109 cells/0.57 mm(2) in the center compared with 1,114 cells/0.57 mm(2) in the controls (P < .05). CD31 expression was greater in the center of the constructs compared with the unseeded scaffolds (P < .05). CONCLUSION: 3D printed ß-TCP and PCL scaffolds seeded with pBMPCs and implanted early into porcine mandibular defects display good bone penetration depth. Further study with a larger sample and larger bone defects should be performed before human applications.

Cyclo-oxygenase-2 expression is associated with vascular endothelial growth factor C expression and lymph node metastasis in oral squamous cell carcinoma.

PURPOSE: Cervical lymph node metastasis in oral squamous cell carcinoma (OSCC) is recognized as a poor prognostic factor, although its mechanism remains unclear. Recently, cyclo-oxygenase-2 (COX-2) level has been found to correlate highly with vascular endothelial growth factor C (VEGF-C) and lymph node metastasis, as in other solid tumors. However, there has been no report of this correlation in OSCC. Therefore, the aim of this study was to investigate whether COX-2 immunohistochemical expression in OSCC was associated with VEGF-C expression, histopathologic parameters, and lymph node metastasis. MATERIALS AND METHODS: Lymphatic vessel density, VEGF-C, and COX-2 immunohistochemical expression were examined pathologically in 60 specimens of invasive OSCC. Relations of histopathologic parameters to lymph node metastasis were analyzed. RESULTS: Expression levels of VEGF-C and COX-2 and lymphatic vessel density in the lymph node metastatic group were significantly higher than in the nonmetastatic group (P < .01). A significant correlation was found between the expression levels of VEGF-C and COX-2 (r = 0.512; P < .001). COX-2 expression was significantly related to lymph node metastasis (P = .004) and VEGF-C expression (P = .005). Univariate analysis showed that survival time was impaired by higher COX-2 and VEGF-C expression levels. Multivariate survival analysis showed that COX-2 expression was an independent prognostic factor. CONCLUSION: This study showed that VEGF-C expression was upregulated by COX-2 in OSCC. High VEGF-C expression appears to promote peritumoral lymphangiogenesis. These data indicated that lymph node metastasis is promoted by COX-2 and VEGF-C in OSCC.

Growth factor directed chondrogenic differentiation of porcine bone marrow-derived progenitor cells.

BACKGROUND: Despite advances in surgical technique, reconstruction of a mandibular condyle still causes significant donor-site morbidity. The purpose of this study was to compare the effect of 3 different growth factors and define optimal cell culture conditions for bone marrow-derived progenitor cells to differentiate into chondrocytes for mandibular condyle reconstruction. METHODS: Porcine bone marrow-derived progenitor cells (pBMPCs) were cultured as a pellet for 2, 3, and 4 weeks under the following conditions: group 1, TGF-ß3 + standard medium; group 2, TGF-ß3 + BMP-2 + standard medium; group 3, TGF-ß3 + IGF-1 + standard medium; and group 4, TGF-ß3 + BMP-2 + IGF-1 + standard medium. Chondrogenic differentiation was evaluated using 3 lineage differentiation markers. RESULTS: The mean type II collagen positive area increased over weeks 2, 3, and 4 in group 4 compared to all the other groups (ANOVA; P = 0.005). At week 4, there was significantly greater type II collagen production in group 4 compared to all the other groups (ANOVA; P = 0.003). The medium in group 4 produces the greatest amount of cartilage when compared to groups 1, 2, and 3, and that 4 weeks produces the greatest amount of type II collagen. CONCLUSIONS: The results of this study indicate that the most efficacious medium for chondrogenic differentiation of pBMPCs was group 4 medium and the most type II collagen was produced at 4 weeks.

Application of custom-made bioresorbable raw particulate hydroxyapatite/poly-L-lactide mesh tray with particulate cellular bone and marrow and platelet-rich plasma for a mandibular defect: evaluation of tray fit and bone quality in a dog model.

The purpose of this study was to evaluate tray fit and bone quality of particulate cancellous bone and marrow (PCBM) mandibular reconstruction using custom-made bioresorbable forged composites of a raw particulate hydroxyapatite/poly-L-lactide (HA/PLLA) tray in a dog model. Mesh sheets of HA/PLLA were formed in a tray shape according to the mandible stereolithographs of 14 beagle dogs. Platelet-rich plasma (PRP) was obtained from venous blood, and PCBM was harvested from the iliac crest. Bone defects were made bilaterally on the lower borders of the mandible. The PCBM and PRP were mixed and compressed into the defects and a custom-made HA/PLLA or a manually adopted titanium tray was fixed by screws. Tray fit and bone qualities were evaluated using computed tomography, microfocus computed tomography and confocal laser scanning microscopy. In buccal side, there is no significant difference with tray fit between the HA/PLLA and the titanium type, but in lingual side, it was better in the HA/PLLA type than that of the Ti type. Bone volume fraction (BV/TV) had markedly increased on the HA/PLLA side at 12 months. In conclusion, the custom-made HA/PLLA tray was easily and accurately adapted to the mandible, and had achieved sufficient bone quality by 12 months.

Three-dimensionally printed polycaprolactone and ß-tricalcium phosphate scaffolds for bone tissue engineering: an in vitro study.

PURPOSE: The purpose of this study was to evaluate porcine bone marrow-derived progenitor cell (pBMPC) proliferation and penetration into a novel 3-dimensionally printed scaffold. MATERIALS AND METHODS: Four different tissue engineering scaffolds to evaluate pBMPC proliferation and penetration were examined. Scaffolds were fabricated from polycaprolactone (PCL) or the combination of ß-tricalcium phosphate (ß-TCP) and PCL (50:50), with 2 separate channel sizes (1 mm [small (S)] vs 2 mm [large (L)]). Scaffolds were fabricated into 20 × 20 × 7-mm blocks by use of a TheriForm machine (Integra Life Sciences, Akron, OH). Four groups of scaffolds were examined for pBMPC proliferation and penetration: group 1, ß-TCP/PCL S; group 2, ß-TCP/PCL L; group 3, PCL S; and group 4, PCL L. Nonparametric mean (Kruskal-Wallis) and multiple comparisons tests were used to compare the 4 groups. RESULTS: No shrinkage or deformation was noted in any of the scaffold groups after 2 weeks of culture. Mean surface cell counts ranged from 13.4 to 87.8 cells/0.57 mm(2), with group 1 (ß-TCP/PCL S) having statistically significantly higher counts than the other groups (P < .001). Mean interior cell counts ranged from 10.9 to 75.6 cells/0.57 mm(2), with group 1 having the greatest interior cell count (P < .001). Total collagen formation ranged from 0.2% to 86%, with group 1 having the highest collagen formation (P < .001). CONCLUSIONS: The 3-dimensionally printed scaffold (ß-TCP/PCL) with 1-mm channels showed greater cellular proliferation, penetration, and collagen formation after a 2-week in vitro culture than the other scaffolds evaluated. ß-TCP/PCL S scaffolds warrant further evaluation for bone tissue engineering in vivo.

Mandibular reconstruction using a tray with particulate cancellous bone and marrow and platelet-rich plasma by an intraoral approach.

PURPOSE: To evaluate the possibility of immediate mandibular reconstruction using particulate cancellous bone and marrow (PCBM), platelet-rich plasma (PRP), and a tray, we compared the postsurgical infection rate and bone formation in patients who received mandibular reconstruction with this method using either an intraoral or extraoral approach. PATIENTS AND METHODS: We conducted a retrospective study of a series of 18 patients who underwent the mandibular reconstruction procedure using a mesh tray with PCBM and PRP, all performed by 1 surgeon. These cases were further divided into those treated by the intraoral approach and those treated by the extraoral approach. Clinical data, postoperative bone formation, and complications in the 2 groups were evaluated. The χ(2) examination and the Mann-Whitney U test were used for statistical analysis. RESULTS: We could not detect any statistically significant differences in clinical data between the 2 groups, except for the timing of reconstruction. There were postoperative complications such as wound dehiscence and tray exposure, as well as infection of the reconstructed bone. The overall complication rate of the recipient sites in the intraoral group was 30% (3 of 10), whereas in the extraoral group, it was 0%. However, satisfactory bone formation was seen in all cases in the intraoral group (100% [10 of 10]) but only 87.5% (7 of 8) in the extraoral group. CONCLUSION: We conclude that mandibular reconstruction using a tray with PCBM and PRP is a safe and reliable method for cases of benign tumor and trauma, even if immediate reconstruction is performed by an intraoral approach.

Clinical application of a custom-made bioresorbable raw particulate hydroxyapatite/poly-L-lactide mesh tray for mandibular reconstruction.

Mandibular reconstruction using particulate cancellous bone and marrow (PCBM) allows functional oral reconstruction. Although ready-made titanium trays are the most common material used in this method, they have some disadvantages such as difficulty in making them form a suitable contour for the defect, and the need for removal. A forged composite of raw particulate hydroxyapatite (HA)/poly-L-lactide (PLLA) is a bioresorbable material that is stronger than pure PLLA and induces bone formation more rapidly. We present two cases successfully treated with custom-made bioresorbable HA/PLLA mesh trays for mandibular reconstruction. A 29-year-old woman with recurrent ameloblastoma and a 66-year-old man with a recurrent keratinized odontogenic tumor of the mandible gave informed consent for this reconstruction technique. Mesh sheets of HA/PLLA were customized by a rapid prototyping method based on computed tomography (CT) data. Marginal resection of the tumor was carried out, and PCBM was harvested from the bilateral posterior iliac crests. PCBM and platelet-rich plasma were transferred to the tray, and the tray was fixed rigidly with HA/PLLA screws. In the second case, dental implants were inserted. There has been no bone resorption for over 2 years since reconstruction in these two cases, and the inserted dental implants have been free from any complications 1 year after loading. The average CT value in Hounsfield units (HU) of the implant sites of two cases was 790. In conclusion, the customized HA/PLLA tray was easily adapted to the mandible, and fine bone quality was obtained. These cases show that this tray system contributed to functional oral rehabilitation with dental implants.

Effect of ibuprofen on osteoblast differentiation of porcine bone marrow-derived progenitor cells.

PURPOSE: Nonsteroidal anti-inflammatory drugs are commonly prescribed to reduce inflammation and pain. However, little is known about the direct effect of these drugs on the differentiation of bone marrow-derived progenitor cells into osteoblasts. The purpose of this study was to determine the effect of ibuprofen on osteoblast differentiation and proliferation in a minipig model. MATERIALS AND METHODS: Bone marrow was aspirated from the minipig ilium, and porcine bone marrow-derived progenitor cells (pBMPCs) were isolated and expanded in standard culture medium. The pBMPCs were replated and differentiated into osteoblasts by use of osteogenic supplements (OS). Five groups were studied: negative control--pBMPCs in standard medium only; positive control--pBMPCs, standard culture medium, and OS; and 3 experimental groups--pBMPCs, standard culture medium, OS, and ibuprofen added in doses of 0.1, 1.0, and 3.0 mmol/L. Cell cultures were evaluated quantitatively by alkaline phosphatase (ALP) stain, von Kossa stain, and deoxyribonucleic acid (DNA) content. RESULTS: pBMPCs cultured with OS and low-dose ibuprofen (0.1 mmol/L) showed ALP stain, von Kossa stain, and DNA content similar to pBMPCs cultured in OS (positive control). pBMPCs cultured in higher doses of ibuprofen (1.0 and 3.0 mmol/L) produced significantly less positive staining of ALP and von Kossa and decreased DNA content. CONCLUSION: The results indicate that high-dose ibuprofen has a deleterious effect on pBMPC differentiation into osteoblasts whereas low-dose ibuprofen does not. The low dose of 0.1 mmol/L is the typical serum level when prescribed for clinical use.

Reconstructing mandibular defects using autologous tissue-engineered tooth and bone constructs.

PURPOSE: Current strategies for jaw reconstruction require multiple operations to replace bone and teeth. To improve on these methods, we investigated simultaneous mandibular and tooth reconstruction, using a Yucatan minipig model. MATERIALS AND METHODS: Tooth and bone constructs were prepared from third molar tooth tissue and iliac-crest bone marrow-derived osteoblasts isolated from, and implanted back into, the same pig as an autologous reconstruction. Implants were harvested after 12 and 20 weeks and evaluated by x-ray, ultrahigh-resolution volume computed tomographic (VCT), histological, and immunohistochemical analyses. RESULTS: Small tooth structures were identified, and consisted of organized dentin, enamel, pulp, and periodontal ligament tissues, surrounded by new bone. No dental tissues formed in implants without tooth-bud cells, and bone regeneration was observed to a limited extent. Immunohistochemical analyses using tooth-specific and bone-specific antibodies confirmed the identity of regenerated tissues. CONCLUSIONS: This pilot study supports the feasibility of tissue-engineering approaches for coordinated autologous tooth and mandible reconstruction, and provides a basis for future improvement of this technique for eventual clinical use in humans.

Tissue engineered hybrid tooth-bone constructs.

Proper rehabilitation of craniofacial defects is challenging because of the complexity of the anatomy and the component tissue types. The ability to simultaneously coordinate the regeneration of multiple tissues would make reconstruction more efficient and might reduce morbidity and improve outcomes. The craniofacial complex is unique because of the presence of teeth, in addition to skin, bone, cartilage, muscle, vascular, and neural tissues since teeth naturally grow in coordination with the craniofacial skeleton, our group developed an autologous, tooth-bone hybrid model to facilitate repair of mandibular defects in the Yucatan minipig. The hybrid tooth-bone construct was prepared by combining tooth bud cell-seeded scaffolds with autologous iliac crest bone marrow derived stem cell-seeded scaffolds, which were transplanted back into surgically created mandibular defects in the same minipig. The constructs were harvested after 12 and 20 weeks of growth. The resulting bone/tooth constructs were evaluated by X-ray, ultra high-resolution volume computed tomography (VCT), histological, immunohistochemical analyses, and transmission electron microscopy (TEM). The observed formation of small tooth-like structures consisting of organized dentin, enamel, pulp, cementum, periodontal ligament, and surrounded by regenerated alveolar bone, suggests the feasibility for regeneration of teeth and associated alveolar bone, in a single procedure. This model provides an accessible method for future clinical applications in humans.

Development of a biodegradable scaffold with interconnected pores by heat fusion and its application to bone tissue engineering.

Tissue engineering has been proposed as an approach to alleviate the shortage of donor tissue and organs by combining cells and a biodegradable scaffold as a temporary extracellular matrix. While numerous scaffold fabrication methods have been proposed, tissue formation is typically limited to the surface of the scaffolds in bone tissue engineering applications due to early calcification on the surface. To improve tissue formation, a novel scaffold with a hierarchical interconnected pore structure on two distinct length scales has been developed. Here we present the fabrication process and the application of the scaffold to bone tissue engineering. Porous poly(lactide-co-glycolide) (PLGA) scaffolds were made by combining solvent casting/particulate leaching with heat fusion. Porcine bone marrow-derived mesenchymal stem cells (MSCs) were differentiated into osteoblasts and cultured on these scaffolds in vitro for 2, 4, and 6 weeks. Subsequently, the constructs were assessed using histology and scanning electron microscopy. The bone marrow-derived osteoblasts attached well on these scaffolds. Cells were observed throughout the scaffolds. These initial results show promise for this scaffold to aid in the regeneration of bone.