Rapid Expansion Protocol Research Articles

Comparison of anti-CD3 and anti-CD28-coated beads with soluble anti-CD3 for expanding human T cells: Differing impact on CD8 T cell phenotype and responsiveness to restimulation

BackgroundThe ability to expand virus- or tumor-specific T cells without damaging their functional capabilities is critical for success adoptive transfer immunotherapy of patients with opportunistic infection or tumor. Careful comparisons can help identify expansion methods better suited for particular clinical settings and identify recurrent deficiencies requiring new innovation.MethodsWe compared the efficacy of magnetic beads coated with anti-CD3 and anti-CD28 (anti-CD3/CD28 beads), and soluble anti-CD3 plus mixed mononuclear cells (designated a rapid expansion protocol or REP) in expanding normal human T cells.ResultsBoth anti-CD3/CD28 beads and soluble anti-CD3 promoted extensive expansion. Beads stimulated greater CD4 cell growth (geometric mean of 56- versus 27-fold (p < 0.01) at day 21) but both stimulated similar CD8 expansion (189- versus 186-fold). Phenotypically, bead-treated CD4 and CD8 T cells and anti-CD3-treated CD4 cells typically assumed an effector/effector memory phenotype by day 14. By comparison, a subset of anti-CD3-treated CD8 cells, derived from naïve cells, retained much greater expression of CD45RA, CD27 and CCR7, than matched bead-treated cells despite comparable expansion. These cells were clearly distinguishable from CD45RA+ terminally differentiated effector cells by the presence of CD27, the absence of CD57 and their inability to produce cytokines after stimulation. When used to expand previously stimulated cells, anti-CD3 plus autologous MNCs produced much less antigen-induced cell death of CD8 cells and significantly more CD8 expansion than beads.ConclusionsAnti-CD3/CD28 beads are highly effective for expanding CD4 cells, but soluble anti-CD3 has significant potential advantages for expanding CD8 T cells, particularly where preservation of phenotypically "young" CD8 cells would be desirable, or where the T cells of interest have been antigen-stimulated in vitro or in vivo in the recent past.

Activation-deactivation rapid palatal expansion and reverse headgear in Class III cases

The aim of this study was to evaluate and compare the dentofacial effects of 1 week rapid palatal expansion (RPE) and activation-deactivation (A/D) RPE protocols with reverse headgear (RH). Two groups, each containing 15 subjects, were included in this study. In the RPE group (seven males and eight females, 11.94 ± 1.62 years), Hyrax screws were activated every 12 hours for 1 week. At the end of this period, RPE was stopped and the patients were instructed to wear the RH. In the A/D-RPE group (seven males and eight females, 11.34 ± 1.81 years), the screws were activated every 12 hours for 1 week. Subsequently, the screws were deactivated every 12 hours for 1 week followed by activation and deactivation for the following 2 weeks. After this protocol, the patients were instructed to use the RH. A total force of 700 g was applied to both groups for 16-18 hours/day for the first 3 months, for 12 hours/day for the second 3 months, and for 6 hours/day for the second 6 months. Lateral cephalometric films were taken before treatment (T1) and at the end of the first (T2) and second (T3) 6 months to evaluate the dentofacial changes. Intragroup differences of each landmark at T2-T1, T3-T2, and T3-T1 were analysed with a paired t-test (P < 0.016), and intergroup differences were compared with an independent t-test (P < 0.05). Anterior movement of point A (4.13 mm) for the A/D-RPE group was approximately twice of the RPE group (2.33 mm; P < 0.001). Backward movement of the mandible showed no significant difference between the groups. Anterior face height increases did not demonstrate significant differences between the groups. The pronounced anterior movement of point A demonstrates that the A/D-RPE procedure positively affects maxillary protraction.

TGF-β1 Induces Preferential Rapid Expansion and Persistence of Tumor Antigen-specific CD8+ T Cells for Adoptive Immunotherapy

Adoptive cell transfer of expanded, autologous tumor-infiltrating lymphocytes (TIL) into lymphodepleted melanoma patients can induce the regression of bulky, metastatic disease. To generate the large numbers of T cells needed for infusion, TIL undergo a rapid expansion protocol (REP) in vitro using anti-CD3 antibody, interleukin-2, and irradiated peripheral blood feeder cells that typically results in an approximately 1000-fold expansion over 14 days. However, we have found that the conventional REP (C-REP) often favors the expansion of CD4+ T cells at the expense of tumor antigen-specific CD8+ T cells, which are the most potent cytolytic effector cells. In this study, we demonstrate that addition of transforming growth factor (TGF)-beta1 to the TIL culture at the onset of rapid expansion (T-REP ) maintained the percentage of CD8+ T cells while not inhibiting overall T-cell expansion. Of T cells expanded from different melanoma patient tumors, 13 of 15 TIL demonstrated improved yields and percentages of both CD8+ and MART-1 melanoma antigen-specific T cells after 14 days of expansion in TGF-beta1 compared with the C-REP. This was associated with a marked improvement in the antitumor activity of the resulting bulk TIL culture in terms of interferon-gamma production and melanoma tumor-specific cytotoxic T-lymphocyte activity. In addition, T-REP T cells demonstrated a higher potential for continued expansion in vitro for up to 3 weeks after the expansion compared with C-REP T cells, suggesting that they may also be capable of increased persistence after adoptive cell transfer. Our results suggest that TGF-beta1-expanded TIL have attributes that might predict efficacy superior to that of conventional TIL.

MART-1–Specific Melanoma Tumor-Infiltrating Lymphocytes Maintaining CD28 Expression Have Improved Survival and Expansion Capability Following Antigenic Restimulation In Vitro

We determined how CD8(+) melanoma tumor-infiltrating lymphocytes (TILs) isolated from two distinct phases of expansion in preparation for adoptive T cell therapy respond to melanoma Ag restimulation. We found that TILs isolated after the rapid expansion protocol (REP) phase, used to generate the final patient TIL infusion product, were hyporesponsive to restimulation with MART-1 peptide-pulsed dendritic cells, with many CD8(+) T cells undergoing apoptosis. Telomere length was shorter post-REP, but of sufficient length to support further cell division. Phenotypic analysis revealed that cell-surface CD28 expression was significantly reduced in post-REP TILs, whereas CD27 levels remained unchanged. Tracking post-REP TIL proliferation by CFSE dilution, as well as sorting for CD8(+)CD28(+) and CD8(+)CD28(-) post-REP subsets, revealed that the few CD28(+) TILs remaining post-REP had superior survival capacity and proliferated after restimulation with MART-1 peptide. An analysis of different supportive cytokine mixtures during the REP found that a combination of IL-15 and IL-21 facilitated comparable expansion of CD8(+) TILs as IL-2, but prevented the loss of CD28 expression with improved responsiveness to antigenic restimulation post-REP. These results suggest that current expansion protocols using IL-2 for melanoma adoptive T cell therapy yields largely CD8(+) T cells unable to persist and divide in vivo following Ag contact. The few CD8(+)CD28(+) T cells that remain may be the only CD8(+) TILs that ultimately survive to repopulate the host and mediate long-term tumor control. A REP protocol using IL-15 and IL-21 may greatly increase the number of CD28(+) TILs capable of long-term persistence.

Dentoskeletal Effects of Maxillary Protraction in Cleft Patients with Repetitive Weekly Protocol of Alternate Rapid Maxillary Expansions and Constrictions

To evaluate the amount of maxillary protraction with face mask in complete unilateral cleft lip and palate patients submitted to two distinct rapid maxillary expansion (RME) protocols. The sample consisted of 20 individuals (nine boys and 11 girls; mean age of 10.4 +/- 2.62 years) with unilateral complete cleft lip and palate who had a constricted maxilla in the vertical and transverse dimensions. Ten patients underwent 1 week of RME with screw activation of one complete turn per day, followed by 23 weeks of maxillary protraction (group 1). The other 10 patients underwent 7 weeks of alternate rapid maxillary expansion and constriction, with one complete turn per day, followed by 17 weeks of maxillary protraction (group 2); both groups underwent a total of 6 months of treatment. Cephalometric measurements were taken at different times: pretreatment (T1), soon after RME (T2), and after 6 months of treatment (T3). Each measurement was analyzed with mixed models for repeated measures, and the covariance structure chosen was compound symmetry. The maxilla displaced slightly forward and downward with a counterclockwise rotation; the mandible rotated downward and backward, resulting in an increase in anterior facial height; the sagittal maxillomandibular relationship was improved; the maxillary molars and incisors were protruded and extruded; and the mandibular incisors were retroclined. There was no significant difference between the groups in evaluation time.

Effect of clinical status and previous treatment of melanoma patients on expansion of TIL for adoptive T-cell therapy

3021 Background: Adoptive cell therapy (ACT) using tumor-infiltrating lymphocytes (TIL) is a promising therapy for the treatment of metastatic melanoma. Trials at the NCI have shown a 50% response rate. However, outside of the NCI, ACT has been quite limited and little is known about how patient characteristics and prior treatment affect large-scale TIL expansion needed for treatment. Methods: As part of a phase II melanoma ACT clinical trial at M. D. Anderson Cancer Center, we evaluated how clinical characteristics such as age, gender, biopsy site, melanoma type, primary tumor location, and prior treatment of patients (N=134) affected TIL expansion rates. 159 isolated metastatic melanomas (19 patients having &gt;1 tumor harvest) were analyzed for the ability to expand a minimum of 40 million TIL, considered a minimum number for the subsequent rapid expansion protocol (REP) needed for TIL infusion. Effects of these parameters on TIL anti-tumor reactivity and TIL/tumor fragment were also analyzed. Statistical analysis was done by either Fisher's Exact test, Wilcoxon Rank Sum test, or Kruskal-Wallis Test. Results: 54.7% of the samples evaluated generated ≥ 40 million TIL, and 70% of 70 samples tested had anti-tumor reactive TIL. There was a significant relationship between age categories (p=0.0216) and gender (p=0.0164) on TIL expansion success. Younger patients did the best and females did better than males (odds 2.19, p=0.044). The site of the primary melanoma affected success of TIL expansion (p=0.0059), while no effect of tumor biopsy site and histological type was found. Prior chemotherapy (or biochemotherapy) greatly reduced TIL growth (odds 0.32, p=0.010) when compared to no prior therapy. Multivariate linear regression analyses showed that prior chemotherapy decreased TIL/tumor fragment yield by 6.19 million (p=0.0004) while immunotherapy increased it by 3.51 million (p=0.16) compared to no prior therapy. Total TIL yield or TIL/tumor fragment did not correlate with patient overall survival. Conclusions: Parameters such as age, sex, primary melanoma site, and prior therapy significantly affect TIL expansion and should be considered in selecting patients for melanoma ACT. No significant financial relationships to disclose.

Single-pass, closed-system rapid expansion of lymphocyte cultures for adoptive cell therapy

Adoptive cell therapy (ACT) for metastatic melanoma involves the ex vivo expansion and re-infusion of tumor infiltrating lymphocytes (TIL) obtained from resected specimens. With an overall objective response rate of 56%, this T-cell immunotherapy provides an appealing alternative to other therapies, including conventional therapies with lower response rates. However, there are significant regulatory and logistical concerns associated with the ex vivo activation and large-scale expansion of these cells. The best current practice uses a rapid expansion protocol (REP) consisting of an ex vivo process that occurs in tissue culture flasks (T-flasks) and gas-permeable bags, utilizes OKT3 (anti-CD3 monoclonal antibody), recombinant human interleukin-2, and irradiated peripheral blood mononuclear cells to initiate rapid lymphocyte growth. A major limitation to the widespread delivery of therapy to large numbers of melanoma patients is the open system in which a REP is initiated. To address this problem, we have investigated the initiation, expansion and harvest at clinical scale of TIL in a closed-system continuous perfusion bioreactor. Each cell product met all safety criteria for patient treatment and by head-to-head comparison had a similar potency and phenotype as cells grown in control T-flasks and gas-permeable bags. However, the currently available bioreactor cassettes were limited in the total cell numbers that could be generated. This bioreactor may simplify the process of the rapid expansion of TIL under stringent regulatory conditions thereby enabling other institutions to pursue this form of ACT.

A phase I trial for the treatment of purine analog-refractory chronic lymphocytic leukemia using autologous T cells genetically targeted to the B cell specific antigen CD19

3045 Background: We have developed a novel immunotherapy strategy based on the genetic modification of patient T cells to recognize the B cell-specific cellular antigen CD19, expressed on B cell tumors, through the retroviral expression of a chimeric antigen receptor (CAR) specific for CD19 (19–28z). Methods: We have initiated a clinical trial utilizing 19–28z+ autologous T cells in patients with purine analog-refractory chronic lymphocytic leukemia (CLL) (IRB #06–138). Enrolled patients initially undergo a leukopheresis procedure in order to obtain T cells. T cells are isolated and activated using Dynabeads ClinExVivo CD3/CD28 magnetic beads. Activated T cells are subsequently transduced with the CD19 specific 19–28z CAR using cGMP gammaretroviral vector stocks generated in our facility, and expanded utilizing a WaveTM bioreactor platform-based rapid expansion protocol. To assess safety, patients enrolled in the first cohort of this trial received an infusion of the lowest planned dose of modified T cells alone, subsequent cohorts will receive infusions of 19–28z+ T cells at the lowest planned dose level following escalating doses of cyclophosphamide chemotherapy followed by escalating doses of T cells after cyclophosphamide. Results: Patients treated in the first cohort with the lowest modified T cell dose alone experienced grade 2 fevers and rigors during infusion but no dose limiting toxicities. Treated patients variably experienced decrease in lymph node size, decreased CD19+ B cell numbers in the peripheral blood, and a decreased dependence on red blood cell transfusions. Conclusions: Infusion of genetically modified T cells targeted to CD19 is well tolerated in patients with refractory CLL with objective evidence of anti-tumor responses. We plan to enroll patients on the second planned cohort of this trial wherein patients will receive prior lymphodepleting chemotherapy with cyclophosphamide. The trial presented here is both the first to utilize gene modified autologous T cells for the treatment of CLL, as well as the first to target CD19+ tumors utilizing a rapid T cell expansion protocol, which represents a promising and novel treatment approach for patients with B cell malignancies. No significant financial relationships to disclose.

Ossificação da sutura palatina mediana após o procedimento de expansão rápida da maxila: estudo radiográfico

OBJETIVO: o presente trabalho tem como objetivo acompanhar radiograficamente a evolução da ossificação da sutura palatina mediana em pacientes submetidos à expansão rápida da maxila, bem como comprovar a validade de se avaliar a neoformação óssea através deste exame complementar de diagnóstico. METODOLOGIA: a amostra constou de 38 pacientes no estágio de dentadura mista que se submeteram ao protocolo de expansão rápida da maxila, sendo 2/4 de volta pela manhã e 2/4 de volta à noite, totalizando 1 volta completa por dia, compreendendo um período de 7 dias. RESULTADOS: radiografias oclusais de maxila tomadas no pós-tratamento evidenciaram uma variação individual quanto ao período necessário para a completa neoformação óssea da sutura palatina mediana. A imagem radiográfica apresentou-se como um método confiável para a determinação da época correta de remoção do aparelho expansor. Tendo em vista a estabilidade pós-tratamento a longo prazo, o aparelho expansor deve ser removido somente após a completa ossificação da sutura. CONCLUSÕES: os resultados revelaram que são necessários mais de três meses para que o processo de ossificação e reorganização sutural se processe após o procedimento de expansão rápida da maxila.

Antigen-independent and antigen-dependent methods to numerically expand CD19-specific CD8 + T cells

Preclinical and clinical trials are investigating the potential of T cells genetically modified to express a first-generation CD19-specific chimeric antigen receptor (CAR), designated CD19R, for adoptive immunotherapy of B-lineage leukemias and lymphomas. Currently, our genetically modified CD19-specific CD8+ (CD19R+CD8+) T cells are expanded ex vivo using a rapid expansion protocol (REP) to clinically meaningful numbers after antigen-independent activation with anti-CD3epsilon and recombinant human interleukin-2 on a double-cell feeder-layer of gamma-irradiated allogeneic peripheral blood mononuclear cells and a lymphoblastoid cell line. We now compare the ability of the REP with CD19-dependent numerical expansion using CD19+ artificial antigen-presenting cells to propagate CD19R+CD8+ T cells. We evaluated long-term (28 days) propagation, CD19R CAR expression, and cytolytic activity of CD19R+CD8+ T cells expanded by either a REP or an antigen expansion protocol (AEP) using K562-derived artificial antigen-presenting cells coexpressing CD19 antigen and two T-cell costimulatory molecules (4-1BB ligand and major histocompatibility class I-related chains A) in the presence of exogenous recombinant human interleukin-2 and recombinant human interleukin-15. Populations of CD19R+CD8+ T cells could be numerically expanded on AEP to meet anticipated clinical need. The AEP was superior to REP, as this method selected for an outgrowth of T cells with increased CD19R CAR expression and improved redirected cytolytic activity. Robust propagation of CD19R+CD8+ T cells achieved by AEP supports qualifying this cell line for use in current good manufacturing practices for CAR+ T cells as an alternative to REP for adoptive immunotherapy clinical trials.

A Novel Strategy for Generation of Human Tumor-Specific T Cell Clones for Adoptive Transfer.

Adoptive T-cell therapy using donor lymphocyte infusions is a promising approach for treating hematological malignancies. But, efficacy is limited by the induction of graft-versus-host disease. Transfer of tumor-specific T-cell clones could enhance the graft-versus-tumor effect and eliminate graft-versus-host disease. However, isolating antigen-specific T-cell clones by the traditional limiting dilution approach is a time-consuming and laborious process. Here, we describe a novel strategy for rapidly cloning tumor-specific T cells. Lymphoma-specific T-cell lines were generated from two follicular lymphoma patients by repeated in vitro stimulation of lymphocytes isolated from tumor or blood with autologous soluble CD40 ligand-activated tumor cells. After four in vitro stimulations at 10-day intervals in the presence of IL-2 and IL-15, T-cell lines were found to be predominantly CD4+ T cells and produced significant amounts of TNF-a, GM-CSF, and IFN-γ in response to autologous tumor cells. The tumor reactivity was MHC class II restricted suggesting that it was mediated by CD4+ T cells. Staining with a TCR Vb antibody panel, a set of monoclonal antibodies against 24 human TCR Vb families, revealed that certain Vb families were overrepresented in each CD4+ T-cell line. In patient 1, 51% of CD4+ T cells were Vb1 positive, and in patient 2, 27% of CD4+ T cells were Vb8 positive. To clone lymphoma-specific T cells, CD4+ T-cell lines were labeled with CFSE and stimulated with autologous tumor cells. After 9 days of in vitro expansion in the presence of IL-2 and IL-15, CD4+ T-cell lines were stained with an anti-human CD4-APC monoclonal antibody and an anti-human TCR Vb-PE monoclonal antibody for each CD4+ T-cell line. Proliferating Vb1 cells from patient 1 and Vb8 cells from patient 2 were identified by their reduction in CFSE staining, and CD4+TCRV b +CFSEdim cells were sorted by flow cytometer. Monoclonality of the sorted cells was confirmed by PCR using a panel of optimized primers specific for 24 TCR Vb families, by TCR Vb spectratype analysis, and finally, by sequencing the TCR Vb gene used by each T-cell clone. Sorted tumor-specific T-cell clones could be expanded to large numbers using a 14-day rapid expansion protocol with allofeeder PBMCs, and confirmed to retain specificity against autologous tumor cells in a cytokine induction assay. This approach was also successfully used to isolate melanoma-specific CD8+ T-cell clones from two patients. We conclude that this approach is highly reproducible, rapid, and efficient for generating antigen-specific T-cell clones for adoptive T-cell therapy against human malignancies in the autologous or allogeneic setting.

Latent-Membrane 2a Specific CTL's but Not Latent-Mebrane 1 Specific CTL's Lysis Efficiently Hodgkin Lymphoma Cells.

Relapse is the leading cause of treatment failure after allogeneic SCT of Hodgkin Disease (HD). As Ebstein-Barr infection (EBV) is associated with 60% of all HD cases, adoptive immunotherapy with donor derived EBV-specific T-cells lines has resulted in disease control of allogeneic SCT. Potential targets for the adoptively transferred T-cells are the type II latency protein LMP-1 and LMP-2a, which are both homogenously expressed by HD cells. In healthy individuals, both LMP-1 and LMP-2a elicits subdominant CD8+ T-cell responses with frequencies of less than 1:10000. LMP-1 and LMP-2a specific T-cells from 1x108 PBMC derived from HLA A*0201+healthy donors were stimulated with the HLA A*0201 LMP1-epitopes YLLEMLWRL and YLGQNWWTL and the HLA A*0201 LMP-2a epitope CLGGLLTM. Activated T-cells were selected by the cytokine secretion assay and expanded for 10 days. In 85% of donors 1.7 x106 (range 0.7 –4.5 x106; n=13) LMP-1 or LMP-2a specific CD8+ T-cell could be generated with an average purity of 83% as determined by tetramer staining. LMP1- and LMP2a-specific CD8+ T-cells were then expanded 3000 x in 14 d by the rapid expansion protocol and evaluated functionally for cytokine production and specific lysis. Both LMP-1 and LMP-2a specific CD8+ T-cells retained specific cytokine production if stimulated with peptide pulsed targets, efficiently lysed peptid pulsed targets. Surprisingly, if LMP-1 was presented endogenously by EBV positive targets or by targets cells transduced with LMP-1, no cytokine production or specific lysis was detected despite protein expression of LMP-1 in all targets. In contrast, IFN-γ production could be readily detected in LMP-2a-specific CD8+ T-cells after stimulation with target cells processing endogenously the LMP-2a antigen as well as specific lysis of EBV positive target cells. Furthermore, LMP2a specific CD8+ demostrated also specific lyse of Hodgkin-cells expressing the LMP2a (30:1 E/T ratio; 29,3%) where as LMP-1-specific CD8+ T-cells could not lyse HD-cells. In summary, LMP-1 and LMP-2a specific T-cells, although present at undectable levels in healthy donors, can be readily selected and expanded to up to 6x109 antigen-specific T-cells in less than 4 weeks starting from 1x108 PBMC. Based on this data, adoptive immunotherapy of relapsed EBV positive HD after allogeneic SCT should be preferentially performed with LMP-2a specific CD8+ T-cells rather than with LMP-1 specific CD8+ T-cells.

Generation of tumor-infiltrating lymphocyte cultures for use in adoptive transfer therapy for melanoma patients.

The generation of T lymphocytes with specific reactivity against tumor antigens is a prerequisite for effective adoptive transfer therapies. Melanoma-specific lymphocyte cultures can be established from tumor infiltrating lymphocytes (TILs) by in vitro culture in high levels of IL-2. We have optimized methods for generating melanoma-reactive TIL cultures from small resected tumor specimens. We report a retrospective analysis of 860 attempted TIL cultures from 90 sequential melanoma biopsy specimens from 62 HLA-A2+ patients. Multiple independent TIL derived from a single tumor often exhibited substantial functional and phenotypic variation. Tumor specific activity was detected in TIL from 29 (81%) of 36 patients screened. TIL cultures selected for high activity were generally capable of large numerical expansion using a single round of a rapid expansion protocol. Limited clonal T-cell populations in an oligoclonal TIL culture could confer specific tumor recognition in these highly selected, highly expanded TIL cultures. These methods were efficient at generating TILs suitable for adoptive transfer therapy.

Proposition of a feasible protocol to evaluate salt sensitivity in a population-based setting.

Although a variety of techniques have been devised to assess salt sensitivity, most have proven cumbersome from a methodological perspective. We therefore attempted to develop a 2-week method by which participants could be tested in an outpatient setting without requirement of a strict dietary regimen. In this method, subjects take 140 mEq of an NaCl supplement per day for 1 week and 25 mg of hydrochlorothiazide daily for another week while maintaining their customary diet. In our first trial, 8 healthy volunteers submitted to this method, as well as to a widely-used rapid volume expansion and contraction protocol. Blood pressure measurements, blood sampling and 24-h urine collection were performed before, in the middle of, and after each intervention. There was a fair correlation (r = 0.69) between the two protocols with respect to the changes in mean blood pressure (deltaMBP), a measure of salt sensitivity. In our second trial, we tested the method on 82 Japanese subjects who had never been treated with antihypertensive drugs. DeltaMBP was significantly correlated with plasma renin activity (PRA) during salt loading (r = 0.52, p < 0.0001) and with the changes in atrial natriuretic peptide (deltaANP) (r = -0.34, p = 0.0018). When total subjects were divided into two subgroups by age, a similar tendency of correlation was observed. Age, PRA during salt loading, deltaANP, and delta norepinephrine were proven to be significant predictors of salt sensitivity and accounted for 46% of the deltaMBP variances. Based on these results, the dietary method presented here seems to be applicable for a population-based survey. Our preliminary data also suggest that PRA and ANP would be of predictive value in the salt sensitivity test.

Continuous versus Conventional Tissue Expansion

Historically, tissue expansion is a prolonged process, typically requiring at least 6 weeks to complete. Recently, interest has increased in shortening this time period. In the current study, a continuous infusion device maintaining constant expander pressure less than capillary filling pressure was used in a canine model in seven dogs to minimize the time period needed to achieve significant expansion. There were no complications, except one device malfunction, corrected by changes in design. The process was shown to be a safe and effective means of producing amounts of expansion similar to traditional methods in approximately 72 hours, with expansion of 28 percent (n = 6) for continuous tissue expansion (CTE) versus 34 percent (n = 6) for a 2-week rapid expansion protocol. This expansion was derived from either stretch of preexisting tissue (46 percent for CTE, 35 percent for 2-week expansion) or recruitment of adjacent tissue. The clinical application of continuous tissue expansion could permit the advantages of tissue expansion to be obtained in many more situations than are currently available to traditional tissue expansion techniques.