Periodic Reporting for period 1 - Laser bioprinting device and in vivo applications (Laser bioprinting device and in vivo applications)
Periodo di rendicontazione: 2023-05-01 al 2024-10-31
PhosPrint's D-LIB platform technology is based on a compact and automated laser bioprinter and protocols for high-speed and high precision in-vivo bioprinting of tissues with autologous human body characteristics. PhosPrint develops the first in-vivo bioprinting tool for bladder tissue repair. We have fully functional industrial laser bioprinter which has been extensively tested, optimized and validated in the surgery room by surgeons, IP (5 patent families) and trade secrets, protocols for cell isolation and expansion, and proprietary pre-clinical data. PhosPrint has currently completed the pre-clinical studies in which bladder tissue has been successfully printed in pigs which have undergone cystectomy during orthotopic neobladder surgery, establishing the safety and efficacy of the approach. Printed neobladders show excellent function 8 weeks post-surgery. The goal of this procedure is to eliminate side-effects associated with the incompatibility of intestinal epithelium with the functional requirements of the urothelium (barrier function, absence of mucus secretion). The outcome is expected to be a hybrid tissue made of smooth muscle lining of intestinal origin and urothelial lining made of autologous urothelial cells which will eliminate metabolic defects, the need for intravesical washings to remove mucus, and the risk for stone formation later on.
Currently, we have two fully functional bioprinters:
One is dedicated to pre-clinical studies and has already been used successfully for that purpose. The second is an upgraded version with the same functionalities, intended for use in first-in-human clinical trials. A second device of this upgraded version will be ready soon to support parallel surgeries at the participating hospitals. We have enhanced our secured IP portfolio from 4 to 5 patent families.
We have developed trade secrets, protocols for cell isolation and expansion, and proprietary pre-clinical data.
We established a Quality Management System (QMS) in collaboration with a Regulatory consulting company based in USA, to ensure compliance with regulatory requirements of both EMA and FDA. Within this QMS system, a design control process was initiated in May 2023 and is currently underway. The primary outcome of Phase I of the QMS implementation was the definition of product requirements, upon which, the design of the clinical device is based.
Each requirement on this list is translated into a specification for the final clinical device, thereby determining the characteristics of the device to comply with MDR745 and the surgical/printing protocol complies with Advanced Therapeutic Medicinal Product requirements produced under GMP requirements. We are currently in Phase 2 of this process.
Key accomplishments WP1:
1. We developed a printer which can print large grafts areas in vivo.
2. A clinical version of this device is now ready for first-in-human trials. Following PhosPrint's QMS design control procedures, the final design and critical functionalities of the device have been successfully established and validated.
3. Additionally, PhosPrint has developed proprietary software to control the printer, ensuring it meets all applicable medical device standards and regulatory requirements. This device and software are now prepared for FIH use.
Key Accomplishments WP 2:
4. Optimization of D-LIB application in pigs
PhosPrint optimized the pig urothelial cell isolation protocol from bladder biopsies. The timing and biospy collection procedure have been fully established. Applying this methodology, fully autologous D-LIB treatment in pigs is realized. Additional improvements and oprimizations in the surgical protocol have been implemented after intensive experimentation in 17 surgeries.
Moreover, we have identified swine cell-compatible antibodies which allow us to histologically characterize cells in culture but also within the D-LIB generated graft upon sacrificing the experimental animals (see below).
Additionally, we have been adapting published protocols and developing own protocols for the expansion of human urothelial cells from healthy human bladder biopsies. Media composition, coating agents and passaging solutions have been tested and optimized for efficient isolation and expansion of human urothelial cells under GMP compliance.
5. Preclinical validation study
A preclinical study was designed and implemented in 12 pigs. Control and D-LIB treated animals were monitored for two months after surgery, and blood and urine samples were also collected at pre-defined time-points. The entire process was continuously monitored and documented complying to our QMS. The preclinical protocol investigated the safety and effectiveness of the D-LIB approach in different drug (cell numbers) doses. The preclinical trial was completed successfully and validated the safety and superiority of D-LIB application to the current gold standard treatment. D-LIB treated animals at 2 months post-surgery, were free from mucus and metabolic defects, in sharp contrast to controls that developed all known side-effects observed in orthotopic neobladder patients.
Key Accomplishments WP 3:
6. Investor update
Significant break-through in our on-going focus and efforts to attract interest from a strategic/lead co-investor that started from day 1 of the project.
We have the commitment of €100k from angel investor, Lars Rasmussen. Greek VC Fund Coralia Ventures VC, following a successful proposal submission and interview, has confirmed their interest and intent to invest in PhosPrint (via email on Friday afternoon) and offered a pre-money valuation of PhosPrint at €4M, to initiate the negotiation and proceed with due diligence.
Additionally, Capital Cell Online investment platform in Healthcare and Biotechnology will be hosting PhosPrint on their Capital Cell funding platform following the successful review from the BioExpert network in February and given that we have the participation from Coralia VC since their policy is to co-invest with another investor. Capital Cell investment can be up to €500K.
One is dedicated to pre-clinical studies and has already been used successfully for that purpose. The second is an upgraded version with the same functionalities, intended for use in first-in-human clinical trials. A second device of this upgraded version will be ready soon to support parallel surgeries at the participating hospitals. We have enhanced our secured IP portfolio from 4 to 5 patent families.
We have developed trade secrets, protocols for cell isolation and expansion, and proprietary pre-clinical data.
We established a Quality Management System (QMS) in collaboration with a Regulatory consulting company based in USA, to ensure compliance with regulatory requirements of both EMA and FDA. Within this QMS system, a design control process was initiated in May 2023 and is currently underway. The primary outcome of Phase I of the QMS implementation was the definition of product requirements, upon which, the design of the clinical device is based.
Each requirement on this list is translated into a specification for the final clinical device, thereby determining the characteristics of the device to comply with MDR745 and the surgical/printing protocol complies with Advanced Therapeutic Medicinal Product requirements produced under GMP requirements. We are currently in Phase 2 of this process.
Key accomplishments WP1:
1. We developed a printer which can print large grafts areas in vivo.
2. A clinical version of this device is now ready for first-in-human trials. Following PhosPrint's QMS design control procedures, the final design and critical functionalities of the device have been successfully established and validated.
3. Additionally, PhosPrint has developed proprietary software to control the printer, ensuring it meets all applicable medical device standards and regulatory requirements. This device and software are now prepared for FIH use.
Key Accomplishments WP 2:
4. Optimization of D-LIB application in pigs
PhosPrint optimized the pig urothelial cell isolation protocol from bladder biopsies. The timing and biospy collection procedure have been fully established. Applying this methodology, fully autologous D-LIB treatment in pigs is realized. Additional improvements and oprimizations in the surgical protocol have been implemented after intensive experimentation in 17 surgeries.
Moreover, we have identified swine cell-compatible antibodies which allow us to histologically characterize cells in culture but also within the D-LIB generated graft upon sacrificing the experimental animals (see below).
Additionally, we have been adapting published protocols and developing own protocols for the expansion of human urothelial cells from healthy human bladder biopsies. Media composition, coating agents and passaging solutions have been tested and optimized for efficient isolation and expansion of human urothelial cells under GMP compliance.
5. Preclinical validation study
A preclinical study was designed and implemented in 12 pigs. Control and D-LIB treated animals were monitored for two months after surgery, and blood and urine samples were also collected at pre-defined time-points. The entire process was continuously monitored and documented complying to our QMS. The preclinical protocol investigated the safety and effectiveness of the D-LIB approach in different drug (cell numbers) doses. The preclinical trial was completed successfully and validated the safety and superiority of D-LIB application to the current gold standard treatment. D-LIB treated animals at 2 months post-surgery, were free from mucus and metabolic defects, in sharp contrast to controls that developed all known side-effects observed in orthotopic neobladder patients.
Key Accomplishments WP 3:
6. Investor update
Significant break-through in our on-going focus and efforts to attract interest from a strategic/lead co-investor that started from day 1 of the project.
We have the commitment of €100k from angel investor, Lars Rasmussen. Greek VC Fund Coralia Ventures VC, following a successful proposal submission and interview, has confirmed their interest and intent to invest in PhosPrint (via email on Friday afternoon) and offered a pre-money valuation of PhosPrint at €4M, to initiate the negotiation and proceed with due diligence.
Additionally, Capital Cell Online investment platform in Healthcare and Biotechnology will be hosting PhosPrint on their Capital Cell funding platform following the successful review from the BioExpert network in February and given that we have the participation from Coralia VC since their policy is to co-invest with another investor. Capital Cell investment can be up to €500K.
We have adapted the bioprinting process to print autologous primary urothelial cells in real time, during surgery on denuded intestinal submucosa. This includes optimization and novel methodologies in the following fields:
1. Biopsy collection process and autologous cell isolation protocol
2. Surgery protocol for neobladder formation by adapting surgery protocols used in humans.
3. Printer modules design and printing protocols that ensure full coverage of denuded intestine and maximum viability of cells
4. Following up on the surgery with the use of imaging techniques to measure neobladder capacity, and biochemical analysis
5. Post-mortem histological evaluation using antibodies against appropriate biomarkers to assess urothelium formation, cell differentiation and tissue stratification.
In parallel, we have developed proprietary protocols for human biopsy processing and urothelial cell expansion and assays that ensure safety and compliance with GMP, also covered by the Phosprint QMS.
PhosPrint aims to develop a platform for in vivo bioprinting of cells for tissue engineering. The route to the market will be the in vivo bioprinting of autologous urothelial cells for orthotopic neobladder construction. Our process development trials yielded a fully functional and validated protocol.
1. Biopsy collection process and autologous cell isolation protocol
2. Surgery protocol for neobladder formation by adapting surgery protocols used in humans.
3. Printer modules design and printing protocols that ensure full coverage of denuded intestine and maximum viability of cells
4. Following up on the surgery with the use of imaging techniques to measure neobladder capacity, and biochemical analysis
5. Post-mortem histological evaluation using antibodies against appropriate biomarkers to assess urothelium formation, cell differentiation and tissue stratification.
In parallel, we have developed proprietary protocols for human biopsy processing and urothelial cell expansion and assays that ensure safety and compliance with GMP, also covered by the Phosprint QMS.
PhosPrint aims to develop a platform for in vivo bioprinting of cells for tissue engineering. The route to the market will be the in vivo bioprinting of autologous urothelial cells for orthotopic neobladder construction. Our process development trials yielded a fully functional and validated protocol.