Rosario Rullo

In vitro bone production using stem cells derived from human dental pulp.

To harvest bone for autologous grafting is a daily problem encountered by craniofacial and oral surgeons. Stem cells derived from human dental pulp are able to differentiate in osteoblasts and are a potential source of autologous bone produced in vitro. The authors describe their preliminary results in this new field with its potential application in craniomaxillofacial surgery. Dental pulp was gently extracted from 34 human permanent teeth (all third molars) of patients 19 to 37 years of age. After they were digested, the cells were selected using a cytometer for c-kit, STRO-1, CD34, CD45, and then for CD44 and RUNX-2. This study, made on a considerable number of cases, provided evidence that dental pulp is extremely rich in stem cells, which were c-kit+/CD34+/STRO-1+/CD45−, capable of differentiation toward several stromal-derived differentiated cells and mainly osteoblasts. These findings, supported by the large number of cases, are of great interest for tissue regeneration, tissue-based clinical therapies, and transplantation.

Recent developments in orofacial cleft genetics.

Nonsyndromic cleft of the lip and/or palate (CLP or orofacial cleft) derives from an embryopathy with consequent failure of the nasal process and/or palatal shelves fusion. This severe birth defect is one of the most common malformations among live births. Nonsyndromic CLP is composed of two separate entities: cleft lip and palate (CL±P) and cleft palate only (CPO). Both have a genetic background, and environmental factors probably disclose these malformations. In CL±P, several loci have been identified, and, in one case, a specific gene has also been found. In CPO, one gene has been identified, but many more are probably involved. Because of the complexity of the genetics of nonsyndromic CLP as a result of the difference between CL±P and CPO, heterogeneity of each group caused by the number of involved genes, type of inheritance, and interaction with environmental factors, we discuss the more sound results obtained with different approaches: epidemiological studies, animal models, human genetic studies, and in vitro studies.

Biological response of human bone marrow mesenchymal stem cells to fluoride‐modified titanium surfaces

OBJECTIVES The aim of the present study was to examine the behaviour of human bone marrow-derived mesenchymal stem cells (BM-MSC) to fluoride-modified grit-blasted (F-TiO) titanium surfaces compared with grit-blasted ones (TiO). MATERIAL AND METHODS Implant surfaces were analysed by atomic force microscopy (AFM) and scanning electron microscopy (SEM). BM-MSC were isolated from healthy donors and grown on the implant surfaces. Cell adhesion and proliferation, type I collagen (Col I) synthesis, osteoblastic differentiation (in terms of alkaline phosphatase activity, osteocalcin synthesis and extracellular matrix mineralization) were assessed. Furthermore, the ability to affect the osteoblastic/osteoclastic balance in terms of osteoprotegerin (OPG) and activator of nuclear factor κ B ligand (RANKL) ratio was investigated. RESULTS F-TiO surface showed higher S(a) values (P<0.05) and the presence of nano-scale structures at the AFM and SEM analysis. Comparable cell morphology and similar adhesion values on both surfaces were detected at early time, whereas higher proliferation values on F-TiO samples were observed at 7 and 10 days. Increased Col I and OPG levels for cells grown on F-TiO were found, whereas RANKL was not detectable in any of the conditioned media. BM-MSC showed a similar expression of early and late osteogenic markers on both TiO and F-TiO surfaces. CONCLUSIONS The results of the present study show that the chemical and micro/nano-scale modifications induced by fluoride treatment of TiO-grit blasted surfaces stimulate the proliferation and the extracellular matrix synthesis by BM-MSC, as well as the increase of OPG synthesis, thus preventing osteoclast activation and differentiation.

Porcine‐Derived Xenograft Combined with a Soft Cortical Membrane versus Extraction Alone for Implant Site Development: A Clinical Study in Humans

BACKGROUND An adequate alveolar crest is essential for implant placement in terms of esthetics and function. The objective of this randomized clinical trial was to compare the preservation of the alveolar ridge dimensions following tooth extraction using porcine-derived xenograft combined with a membrane versus extraction-alone (EXT) sites. METHODS Fifteen patients who required double extraction of contralateral premolars and delayed implant placement were randomly selected to receive both ridge-preservation procedure and EXT. The test sites (alveolar ridge preservation [ARP]) included 15 sockets treated using a corticocancellous porcine bone xenograft (OsteoBiol® Gen-Os; Tecnoss srl, Giaveno, Italy) associated with a soft cortical membrane (OsteoBiol® Lamina; Tecnoss srl), while the corresponding control sites (EXT) were left without grafting for EXT. Horizontal and vertical ridge dimensions were recorded at baseline and 6 months after extractions. RESULTS After 6 months, the EXT sites showed a significantly greater reabsorption of the buccolingual/palatal dimension of the alveolar ridge (3.7 ± 1.2 mm) compared with the ARP sites (1.8 ± 1.3 mm). The mean vertical ridge height reduction in the control sockets was 3.1 ± 1.3 mm at the buccal sites and 2.4 ± 1.6 mm at the lingual sites compared with 0.6 ± 1.4 and 0.5 ± 1.3 mm, respectively, in the test sockets. The differences between test and control sockets were not significant for the mesial and distal measurements. CONCLUSIONS The placement of a porcine xenograft with a membrane in an extraction socket can be used to reduce the hard tissue reabsorption after tooth extraction compared with EXT.

A comparison of salivary substitutes versus a natural sialogogue (citric acid) in patients complaining of dry mouth as an adverse drug reaction: a clinical, randomized controlled study

OBJECTIVE We aimed to compare the efficacy of saliva substitutes and citric acid long-term therapy for oral dryness relief and unstimulated salivary flow in patients reporting drug-induced xerostomia. STUDY DESIGN Fifty-four patients reporting drug-induced xerostomia were randomly subdivided into 3 groups and respectively administered artificial saliva, 3% citric acid, or distilled water in mouthwash 4 times a day for 30 days. Patients underwent measurement of unstimulated whole saliva before and after they finished therapy and were asked to note in a daily diary any symptomatologic changes 15 minutes and 1 hour after each daily intake of test solution. RESULTS Fifteen minutes after solution intake, 12 patients (67%) belonging to the artificial saliva group, 9 (50%) from the citric acid group, and 2 (11%) from the water group reported significant symptomatologic improvement. One hour after solution intake, 7 patients (39%) from the artificial saliva group, 10 (56%) from the citric acid group, and 0 from the water group noted significant symptomatologic improvement. None of the drugs tested affected unstimulated whole saliva flow. CONCLUSIONS Both artificial saliva and citric acid provided immediate relief from oral dryness. Citric acid also provided a longer-lasting feeling of oral moistness at 1 hour after use owing to its protracted activity on salivary gland function.

Speech assessment in cleft palate patients : A descriptive study

OBJECTIVE Speech disorders could affect the intelligibility, but also social competence and emotional development of a cleft-palate child. In this study, we proposed to examine the phono-articulatory defects of a group of surgically treated cleft palate patients, relatively to the familial social background, and children ability of self-controlling during spontaneous language. METHODS Sixty-eight children (22 males and 46 females) mean age 6.87 years, affected by a non-syndromic isolated cleft of the palate underwent to the same surgical protocol. Linguistic level was evaluated using the speech assessment protocol of Accordi and colleagues that focuses on intelligibility, nasality, nasal air escape, pharyngeal friction, and glottal stop. Each of these parameters is evaluated on a four-point scale, ranging from 0 to 3. Spearman rank order correlation was used to detect significant association between analyzed parameters; also the patient social background was considered for statistical analysis. RESULTS The 5.88% of the sample had a negative result, with a severe to moderate degree of nasality and nasal air escape, presence of pharyngeal friction and glottal stop and, obviously a systematic compromising of the intelligibility. Also grimace was evident. The 38.23% of the sample showed an acceptable phonological outline; nasality and nasal air escape were mild, and the intelligibility resulted normal. Thirty-eight children (55.88%) showed a perfectly normal speech. Statistical analysis (Spearman rank order correlation) revealed a positive correlation between the severity of nasality and nasal air escape and the patient social contest of life (p</=0.05); moreover, there was a positive correlation between nasality and nasal air escape with the final intelligibility (p<or=0.05), and between the intelligibility and the patient social contest of life. CONCLUSION Phonetic and phonological development in cleft child are not only due to the surgical strategies and the surgeon experience, but also influenced by the willingness to collaborate of the patient and especially of the parents, the timeliness of the logopaedic intervention, and by inborn capabilities of the child to control the emission of the air from nasal and oral cavities.

Interplay between steroid receptors and neoplastic progression in sarcoma tumors

Steroid hormones are expressed at low levels in mesenchymal cells and are highly expressed in soft tissue sarcoma. In human soft tissue fibrosarcoma cell line (HT‐1080), the epidermal growth factor (EGF) stimulates the express of matrix metal (MMPs) expression through a Src‐dependent mechanism. In human fibrosarcomas, increased expression of MMPs correlates with the metastatic progression. Our recent data in human breast cancer cell line MCF‐7, demonstrates that EGF stimulates estradiol receptor (ER) phosphorylation on tyrosine at position 537 thereby promoting the association of a complex among EGF receptor (EGFR), androgen receptor (AR), ER, and Src that activates EGF‐dependent signaling pathway. In the present study, we demonstrate that, in HT‐1080 cells, the Src kinase activity is involved in EGFR phosphorylation and this activity is regulated by an interplay between Src, steroid receptors, and EGFR. In these cells, estradiol (E2)/ER and synthetic androgen (R1881)/AR trans‐activate EGFR leading to the downstream signaling and to ERK activation. Indeed, the association between ER/AR and EGFR enhances metastatic progression of fibrosarcoma tumors. A population pilot study performed on 16 patients with soft tissue neoplasias highlights that MMPs expression correlates with progression of anaplastic sarcoma as well as overexpression of EGFR. These findings suggest that there is a crosstalk among AR, ER, and EGFR that lead to src activation also in fibrosarcoma cells. J. Cell. Physiol. 226: 2997–3003, 2011.

Study of folate receptor genes in nonsyndromic familial and sporadic cleft lip with or without clef palate cases

Folate receptor family members (FOLRs) mediate the delivery of 5‐methyltetrahydrofolate to the interior of, out of within, or between cells in a process known as potocytosis. Three FOLRs and a pseudogene map to 11q13.4. The aim of this study was to verify whether FOLRs could be responsible for the onset of nonsyndromic cleft lip with or without cleft palate (CL/P). Linkage and linkage disequilibrium between genetic markers and disorder were analyzed. Patients and their mothers from 71 familial CL/P pedigrees and 75 sporadic cases from Italian population were investigated by PCR‐SSCP analysis. Data from mutation scanning allowed us to find only a silent mutation in FOLR1 present in a mother and her child. Our findings do not support FOLR1 and FOLR2 genes in the onset of CL/P.

Oral mucosa optical biopsy by a novel handheld fluorescent confocal microscope specifically developed: technologic improvements and future prospects.

OBJECTIVE This pilot study evaluated the baseline effectiveness of a novel handheld fluorescent confocal microscope (FCM) specifically developed for oral mucosa imaging and compared the results with the literature. STUDY DESIGN Four different oral sites (covering the mucosa of the lip and of the ventral tongue, the masticatory mucosa of the gingiva, and the specialized mucosa of the dorsal tongue) in 6 healthy nonsmokers were imaged by an FCM made up of a confocal fiberoptic probe ergonomically designed for in vivo oral examination, using light at the wavelength of 457 nm able to excite the fluorophore acriflavine hydrochloride, topically administered. In total, 24 mucosal areas were examined. RESULTS The FCM was able to distinctly define epithelial cells, bacterial plaque, and inflammatory cells and to image submucosal structures by detecting their intrinsic fluorescence. CONCLUSIONS When compared with other devices, this FCM allowed the user to image each oral site at higher magnification, thus resulting in a clearer view.

The Use of Piezosurgery in Genioplasty.

AbstractThe aim of this study was to compare the postoperative complications during genioplasty using piezosurgery devices or traditional rotating drills. Forty patients underwent genioplasty to solve their chin anomalies. This study showed that, using piezosurgery, pain and swelling were reduced compared to the traditional technique in the immediate postoperative period of healing (within 15 days). Paresthesia was observed in all patients up to 15th postoperative day, independently of the used surgical technique, but at the sixth postoperative month it was completely absent.