Improved in Vitro Generation of Epithelial Grafts With Oral Mucosa

Tomson, A. M.; Scholma, Janny; Blaauw, Eh; Rosingh, H. J.; Dikkers, Frederik G.

doi:10.1097/00007890-199412150-00028

Cited by 3 publications

(2 citation statements)

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“…The harvested buccal mucosa was disinfected with 70% alcohol for 30 s, and then rinsed with phosphate‐buffered saline (PBS) several times to remove any blood, mucous, and alcohol. We followed the culture methods of Thomson et al (14) with the exception of supplement such as 10% fetal bovine serum (FBS, BioWhittaker, Baltimore, MD, U.S.A.) instead of 5% FBS. After washing with PBS, the mucosa was treated with Dispase II (1 mg/mL; Boehringer Mannheim, Germany) at 4°C for 14–16 h to separate the epidermis from the dermis.…”

Section: Methodsmentioning

confidence: 99%

Fabrication of Myomucosal Flap Using Tissue‐engineered Bioartificial Mucosa Constructed With Oral Keratinocytes Cultured on Amniotic Membrane

et al. 2006

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The purpose of this study was to fabricate bioartificial mucosa using cultured oral keratinocytes (OKCs) on an amniotic membrane (AM), and to evaluate the possibility of developing a prelaminated myomucosal flap using the fabricated bioartificial mucosa and local muscle flap. Buccal mucosa was harvested from male New Zealand rabbits (n = 40, 2.5-3.0 kg) and primary cultivation was performed. The cultured OKCs were seeded on the AM and a submerged culture was performed. Prelamination of the bioartificial mucosa was performed on the latissimus dorsi (LD) muscle of rabbits. Survival rate, layer of OKCs, and Cinamon's score (CS) based on macroscopic and microscopic examinations were evaluated 7, 10, 14, and 21 days after prelamination (n = 10 per day). The OKCs cultured on AM showed multiple layers (3.85 +/- 1.32) and cells were tightly adhered with desmosomes. Basal layer cells adhered to the AM with hemidesmosomes. In addition, the AM played an excellent role as a substrate for the OKCs and simplified handling during prelamination. A myomucosal flap with OKCs cultured on AM was fabricated within 2 weeks (CS: 11.05 +/- 2.63). The basement component of laminin was observed 2 weeks after prelamination and showed enough strength to adhere to the underlying fascia. A myomucosal flap was successfully developed using prelamination of bioartificial mucosa on the LD muscle between 10 and 14 days.

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Section: Methodsmentioning

confidence: 99%

Fabrication of Myomucosal Flap Using Tissue‐engineered Bioartificial Mucosa Constructed With Oral Keratinocytes Cultured on Amniotic Membrane

et al. 2006

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“…From GABEB patient 1 a punch biopsy (diameter 6 mm) of involved skin of the upper arm was taken. Epidermal cells were cultured according to a modified procedure of Rheinwald and Green (30). Briefly, epidermal cells were dissociated from the tissue specimen by a 30-min incubation with trypsin and grown in two 25-cm2 culture flasks on a monolayer of lethally irradiated 3T3-mouse fibroblasts in keratinocytes medium (DME-F12/HAM 2:1, 20% FCS, 0.4 Mg/ml hydrocortisone, 10 ng/ml EGF, 10 ,ug/ml non-essential amino acids and 10`-0 choleratoxin).…”

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confidence: 99%

180-kD bullous pemphigoid antigen (BP180) is deficient in generalized atrophic benign epidermolysis bullosa.

Jonkman¹,

Jong²,

Heeres³

et al. 1995

J. Clin. Invest.

163

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IntroductionGeneralized atrophic benign epidermolysis bullosa (GA-BEB) is a form of nonlethal junctional epidermolysis bullosa characterized by universal alopecia and atrophy of the skin. We report a deficiency of the 180-kD bullous pemphigoid antigen in three patients with GABEB from unrelated families. We screened specimens of clinically normal skin from nine junctional epidermolysis bullosa patients (3 GABEB, 4 lethal, 1 cicatricial, 1 pretibial) by immunofluorescence using monoclonal antibodies to the 180-kD and 230-kD bullous pemphigoid antigens (BP180 and BP230). In the skin of the three GABEB patients there was no reactivity with antibodies to BP180, whereas staining for BP230 was normal. In the skin of the other six, non-GABEB patients, included in this study the expression of BP180 and BP230 was normal. Immunoblot analysis of cultured keratinocytes from one of the GABEB patients also failed to detect BP180 antigen, whereas BP230 was present in normal amounts. In previous studies on the expression of the bullous pemphigoid antigen in JEB, sera from bullous pemphigoid (BP) patients were used, the specificity of which had not been analyzed by immunoblotting (6-9). The expression was found to be normal or variable. Later it became apparent that BP-serum is a mixture of polyclonal antibodies, which are very difficult to characterize, even after affinity-purification (10). Fortunately, reliable monospecific antibodies for each of the two major bullous pemphigoid antigens have now become available (1 1, 12).In this study we used monoclonal antibodies specific for BPI 80 and BP230 (11,12). In addition we studied the expression of the newly described 500-kD hemidesmosomal plaque protein HD1 (13)

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Optimal management of the 3-to 6-centimeter anterior urethral stricture

Zinman

2000

Curr Urol Rep

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The optimal management of anterior urethral stricture that does not respond to an endoscopic urethrotomy or is found to be unsuitable for excision and anastomotic repair remains controversial. Genital skin island onlay flaps or buccal mucosal grafts are presently the most dependable single stage procedures used for strictures more than 3 cm in length. Nonhirsute penile island fasciocutaneous flaps constitute the most durable substitution technique for pendulous stricture disease, with long-term studies reporting 90% to 96% success. The complex proximal bulb and bulbomenbraneous stricture with a compromised proximal fibrous avascular bed is ideally managed with either a penile or scrotal island flap or some combination of partial urethral excision with a dorsally placed genital skin island. The buccal mucosal onlay graft is a promising addition to this reconstructive paradigm, and early outcomes have been favorable. The graft is presently used for bulbar strictures, avoiding the transsphincteric on pendulous location, or a compromised recipient bed. The present standard of care for proximal bulb strictures is wide bulbospongiosal mobilization, partial urethral excision, a floor strip anastomosis, and placement of an augmenting flap on the graft in a dorsal location.

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Improved in Vitro Generation of Epithelial Grafts With Oral Mucosa

Cited by 3 publications

References 0 publications

Fabrication of Myomucosal Flap Using Tissue‐engineered Bioartificial Mucosa Constructed With Oral Keratinocytes Cultured on Amniotic Membrane

Fabrication of Myomucosal Flap Using Tissue‐engineered Bioartificial Mucosa Constructed With Oral Keratinocytes Cultured on Amniotic Membrane

180-kD bullous pemphigoid antigen (BP180) is deficient in generalized atrophic benign epidermolysis bullosa.

Optimal management of the 3-to 6-centimeter anterior urethral stricture

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