Lipid-polymer hybrid NPs were prepared via self-assembly of PLGA (poly (D,L-lactic-co-glycolic acid); Lactel, Pelham, AL), lecithin (soybean, refined, molecular weight: ~330 Da; Alfa Aesar, Ward Hill, MA), and DSPE-PEG (1,2-distearoyl-sn-glycero-3-phosphoethanolamine- N-carboxy (polyethylene glycol)2000); Avanti, Alabaster, AL) through a single-step nanoprecipitation method. Briefly, PLGA polymer was dissolved in acetonitrile with concentrations ranging from 1~5 mg/mL. Lecithin/DSPE-PEG (8.5/1.5, molar ratio) with a weight ratio of 15% to the PLGA polymer were dissolved in 4 wt% ethanol aqueous solution. The lecithin/DSPE-PEG solution was heated to 65°C to ensure all lipids were in liquid phase. The resulting PLGA solution was then added into the preheated lipid solution dropwise under gentle stirring. The mixed solution was vortexed vigorously for 3 minutes followed by gentle stirring for 2 hours at room temperature. The remaining organic solvent and free molecules were removed by washing the NP solution three times using an Amicon Ultra-4 centrifugal filter (Millipore, Billerica, MA) with a molecular weight cut-off of 10,000 Da. To prepare drug-encapsulated NPs, docetaxel (Sigma-Aldrich, St Louis, MO) with proper initial dosage was dissolved into the PLGA acetonitrile solution before the nanoprecipitation process. NP size (diameter, nm) and surface charge (zeta potential, mV) were obtained from three repeat measurements by Quasi-elastic laser light scattering with a ZetaPALS dynamic light scattering detector (15 mW laser, incident beam = 676 nm; Brookhaven Instruments Corporation, Holtsville, NY).
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Poly-L-lactic acid
Poly-L-lactic acid
Poly-L-lactic acid (PLLA) is a biodegradable, biocompatible polymer derived from renewable resources.
It has a wide range of applications in biomedical and engineering fields, such as tissue engineering scaffolds, drug delivery systems, and renewable packaging materials.
PLLA exhibits good mechanical properties, slow degradation kinetics, and the ability to be processed into various forms.
Researchers can utilize PubCompare.ai's AI-driven protocol comparision tool to quickly identify the best reproducibble PLLA research protocols from literature, pre-prints, and patents, enhacing efficiency and reproducibility in their studies.
It has a wide range of applications in biomedical and engineering fields, such as tissue engineering scaffolds, drug delivery systems, and renewable packaging materials.
PLLA exhibits good mechanical properties, slow degradation kinetics, and the ability to be processed into various forms.
Researchers can utilize PubCompare.ai's AI-driven protocol comparision tool to quickly identify the best reproducibble PLLA research protocols from literature, pre-prints, and patents, enhacing efficiency and reproducibility in their studies.
Most cited protocols related to «Poly-L-lactic acid»
1,2-distearoylphosphatidylethanolamine
acetonitrile
Alabaster
DA10
Docetaxel
Ethanol
glycolic acid
Hybrids
Lecithin
Lipids
Molar
Pharmaceutical Preparations
Phosphatidylethanolamines
Poly A
polyethylene glycol 2000
Polylactic Acid-Polyglycolic Acid Copolymer
Polymers
Solvents
Soybeans
100 nm polymeric cores were prepared using 0.67 dL g−1 carboxyl-terminated 50:50 poly(lactic-co-glycolic) acid (PLGA) (LACTEL Absorbable Polymers) in a nanoprecipitation process. 1 mL of 10 mg mL−1 PLGA solution in acetone was added dropwise to 3 mL of water. For fluorescently labeled nanoformulations, 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindodicarbocyanine perchlorate (DiD, ex = 644 nm/em = 665 nm, Life Technologies) was loaded into the polymeric cores at 0.1 wt%. The mixture was then stirred in open air for 1 h and placed in vacuum for another 3 h. The resulting nanoparticle solution was filtered with 10 kDa MWCO Amicon Ultra-4 Centrifugal Filters (Millipore). Platelet membrane cloaking was then accomplished by dispersing and fusing platelet membrane vesicles with PLGA particles via sonication using an FS30D bath sonicator at a frequency of 42 kHz and a power of 100 W for 2 min. The size and the surface zeta potential of the replicate PNP samples (n=3) were obtained by DLS measurements using a Malvern ZEN 3600 Zetasizer. PBS stability was examined by mixing 1 mg mL−1 of PNPs in water with 2X PBS at a 1:1 volume ratio. Storability of PNPs was examined by suspending PNPs in 10% sucrose. The nanoparticle solutions were subject to either a freeze-thaw cycle or lyophilization followed by resuspension. The resulting particle solution was then monitored for particle size using DLS. The structure of PNPs was examined with TEM following negative staining with 1 wt% uranyl acetate using an FEI 200 kV Sphera microscope. RBCNPs were prepared using the same polymeric cores and RBC membranes of equivalent total surface area to the platelet membranes using a previously described protocol16 (link). The RBCNPs were characterized using DLS and had similar size and zeta potential as the PNPs.
Docetaxel-loaded PLGA nanoparticle cores were prepared via a nanoprecipitation process. 10 wt% docetaxel was added to 5 mg PLGA in acetone and precipitated dropwise into 3 mL water. The solvent was evaporated as described above and free docetaxel was removed via repeated wash steps. Vancomycin-loaded nanoparticles were synthesized using a double emulsion process. The inner aqueous phase consisted of 25 µL of vancomycin (Sigma Aldrich) dissolved in 1 M NaOH at 200 mg mL−1. The outer phase consisted of 500 µL of PLGA polymer dissolved in dichloromethane at 50 mg mL−1. The first emulsion was formed via sonication at 70% power pulsed (2 sec on/1 sec off) for 2 min on a Fisher Scientific 150E Sonic Dismembrator. The resulting emulsion was then emulsified in aqueous solution under the same dispersion setting. The final w/o/w emulsion was added to 10 mL of water and the solvent was evaporated in a fume food under gentle stirring for 3 h. The particles were collected via centrifugation at 80,000 × g in a Beckman Coulter Optima L-90K Ultracentrifuge. The particles were washed and resuspended in water. Upon preparation of drug-loaded PLGA cores, cell membrane coating was performed by adding the appropriate surface area equivalent of either platelet or RBC membrane followed by 3 min of sonication in a Fisher Scientific FS30D Bath Sonicator. Particle size, polydispersity (PDI), and surface zeta potential were characterized using DLS. Drug loading yield and release rate of replicate samples (n=3) were quantified by high performance liquid chromatography (HPLC). Drug release was determined by dialyzing 500 µL of particle solution at a concentration of 2.67 mg mL−1 in PBS using 3.5K MWCO Slide-A-Lyzers (Thermo Scientific).
Docetaxel-loaded PLGA nanoparticle cores were prepared via a nanoprecipitation process. 10 wt% docetaxel was added to 5 mg PLGA in acetone and precipitated dropwise into 3 mL water. The solvent was evaporated as described above and free docetaxel was removed via repeated wash steps. Vancomycin-loaded nanoparticles were synthesized using a double emulsion process. The inner aqueous phase consisted of 25 µL of vancomycin (Sigma Aldrich) dissolved in 1 M NaOH at 200 mg mL−1. The outer phase consisted of 500 µL of PLGA polymer dissolved in dichloromethane at 50 mg mL−1. The first emulsion was formed via sonication at 70% power pulsed (2 sec on/1 sec off) for 2 min on a Fisher Scientific 150E Sonic Dismembrator. The resulting emulsion was then emulsified in aqueous solution under the same dispersion setting. The final w/o/w emulsion was added to 10 mL of water and the solvent was evaporated in a fume food under gentle stirring for 3 h. The particles were collected via centrifugation at 80,000 × g in a Beckman Coulter Optima L-90K Ultracentrifuge. The particles were washed and resuspended in water. Upon preparation of drug-loaded PLGA cores, cell membrane coating was performed by adding the appropriate surface area equivalent of either platelet or RBC membrane followed by 3 min of sonication in a Fisher Scientific FS30D Bath Sonicator. Particle size, polydispersity (PDI), and surface zeta potential were characterized using DLS. Drug loading yield and release rate of replicate samples (n=3) were quantified by high performance liquid chromatography (HPLC). Drug release was determined by dialyzing 500 µL of particle solution at a concentration of 2.67 mg mL−1 in PBS using 3.5K MWCO Slide-A-Lyzers (Thermo Scientific).
Aluminum
Fibrosis
Gamma Rays
glycolic acid
hexafluoroisopropanol
Molar
Needles
Poly A
Polylactic Acid-Polyglycolic Acid Copolymer
Steel
Viscosity
ITR was purchased from Beta-Pharma (Shanghai, China). Poly(d ,l lactic-co-glycolic acid; PLGA) (50/50, M.W. 45–70 kDa), medium molecular weight chitosan (CS), and tri polyphosphate (TPP) were purchased from Sigma-Aldrich, St. Louis, MO, USA. Β-actin (NB600-50155), Bax (NB100-609655), and Bcl2 (NB100-56098) primary antibodies were purchased from Novus Biologicals (Centennial, CO, USA). The apoptosis detection kit (556547) was purchased from BD Bioscience (NJ, USA). All other chemicals used were of analytical grade.
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Actins
Antibodies
Apoptosis
BCL2 protein, human
Biological Factors
Chitosan
glycolic acid
Novus
Poly A
Polylactic Acid-Polyglycolic Acid Copolymer
Polyphosphates
Most recents protocols related to «Poly-L-lactic acid»
1 g of PLLA was dissolved
in 3 mL of dichloromethane using an ultrasonic bath. Next, 1 g of
ammonium bicarbonate was added. The solution was drop-cast on the
electrode surface, and after 5 min the electrode was dipped in 85
°C for 5 min to initiate pore formation and in cold water for
20 min to quench the reaction. Electrodes were left to dry at room
temperature.27 (link)
in 3 mL of dichloromethane using an ultrasonic bath. Next, 1 g of
ammonium bicarbonate was added. The solution was drop-cast on the
electrode surface, and after 5 min the electrode was dipped in 85
°C for 5 min to initiate pore formation and in cold water for
20 min to quench the reaction. Electrodes were left to dry at room
temperature.27 (link)
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Example 1
In order to prepare a PLLA mixture, first, PLLA (poly-L-lactic acid) having a molecular weight of about 150,000 kDa (kilodaltons) was prepared. CMC (carboxymethylcellulose) and mannitol were mixed with the PLLA, followed by freeze-drying. At this time, the freeze-drying was carried out by primary freeze-drying at −60 to −100° C. for 12 to 24 hours and secondary drying at 15 to 25° C. for 5 to 10 days.
The freeze-dried PLLA mixture was pulverized to a size ranging from 30 μm to 100 μm (appropriately 50 μm) using an overhead stirrer, and was subjected to gamma-ray sterilization to prepare a PLLA mixture powder.
Example 2
A multiple emulsion of water in oil in water (W/O/W) was prepared by the following two-step procedure.
A 5 mM phosphate buffer solution in which a drug containing a pain reliever and/or antibiotic is dissolved in a concentration of 5 mg/ml was mixed with a primary core material (W1), the PLLA mixture prepared in Example 1, a biodegradable polymer, an MCT (medium-chain triglyceride) oil and PGPR (polyglycerol polyricinoleate) as an emulsifier, followed by stirring to obtain an oily phase (O), and the oily phase was homogenized to prepare a W1/O emulsion as a primary emulsion.
In order to prepare a W/O/W emulsion, a secondary wall material (W2) for encapsulating the W1/O emulsion was prepared as follows.
First, HA having a molecular weight of about 2 million kDa was mixed with BDDE (butanediol diglycidyl ether) as a crosslinking agent at a predetermined ratio, and the gelled HA was washed with a phosphate buffer. The phosphate buffer was used in an amount of 30 to 80 liters (appropriately 50 liters) with respect to 100 grams of HA.
The washed HA was passed through a screen with 80 to 120 mesh having a uniform size to obtain crosslinked HA with uniform particles. In this case, the optimal screen hole size may be 100 mesh.
Distilled water was added to the crosslinked HA and was then mixed at 9,400 rpm using a homogenizer for 5 minutes. This material was further homogenized at 14,000 rpm using a homogenizer for 5 minutes to obtain a secondary wall material (W2).
Then, 25% (w/w) of the W1/O emulsion was mixed with 75% (w/w) of the secondary wall material (W2), followed by stirring at 400 rpm in a stirrer for 5 minutes and homogenization using a homogenizer (5 minutes, 20,000 rpm) to prepare a PLLA-HA W/O/W emulsion.
Example 3
A microcapsule as a powder was prepared from the W/O/W emulsion prepared in Example 2 using a spray dryer (Eyela spray-dryer SD-1000, Eyela, Tokyo, Japan). Specifically, the temperature of the fed air was adjusted to 130±5° C., the discharge air temperature was adjusted to 80±5° C., the rotary sprayer was adjusted to 10×10 kPa, the blower speed was adjusted to 0.80 m3/min, and the pump speed was adjusted to 1.0 mL/min.
The double microcapsule prepared through the above process was injected into a vial and then gamma-ray sterilized once more and frozen at −20° C.
The present invention is characterized in that the double microencapsulated PLLA-HA W/O/W emulsion can be used immediately after injecting injection water into the same. Specifically, when preparing a filler for the face, 15 to 25 ml of distilled water was mixed with 10 mg of the double microcapsule, whereas when preparing a filler for the body, to 35 cc of distilled water was mixed with 10 mg of double microcapsule.
The double microencapsulated PLLA-HA W/O/W emulsion was completely micronized and homogenized during the preparation process, thereby solving conventional problems in which it is required to form a suspension and allow the suspension to stand for 2 hours or longer before use after mixing water for injection, and particles of PLLA agglomerate in the composition.
As described above, the present invention is characterized in that the long time taken to form initial volume during injection, which is the disadvantage of conventional PLLA filler products (e.g., products such as Sculptra), can be shortened by 6-8 weeks or more, and the present invention is based on a combination with cross-linked hyaluronic acid in order to minimize the formation of granuloma, which is another disadvantage of conventional PLLA fillers, and the aggregation of PLLA is significantly reduced even after insufficient mixing time or long-term storage by producing the same into fine particles.
Further, the sustained-release double microcapsule of the present invention is in a form in which a pain reliever or antibiotic is contained in PLLA as a biodegradable polymer particle, and is characterized by continuously releasing the pain reliever as it is decomposed in the body, thereby solving problems related to pain and infection that may be caused by filler treatment.
The description of the present invention is provided only for illustration, and those skilled in the art will appreciate that various modifications, additions and substitutions are possible, without departing from the scope and spirit of the invention as disclosed in the accompanying claims. Therefore, it should be understood that the embodiments described above are illustrative and non-limiting in all respects. For example, a component described as a single unit may be implemented in a separated manner, and similarly, components described as being separated may also be implemented in a combined form.
The scope of the present invention is defined by the claims to be set forth below, and all alterations or modifications derived from the meanings and scopes of the claims and equivalents thereto should be construed as falling within the scope of the present invention.
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All reagents and solvents were of analytical grade. The PLGA (Purasorb® PDLG 5002A, Corbion N.V.) NPs were prepared with minor modifications, as previously reported [35 (link)]. All types of PLGA NPs were purchased from Nanoglia (Daejeon, Republic of Korea) [25 (link)]. To prepare the siRNA-encapsulated PLGA NPs, 20 μmol of each siRNA (Invitrogen; Ca# 53,640, The sequence of p16ink4a siRNA as follow; sense 5’-GGUGAUGAUGAUGGGCAACtt-3’, antisense 5’-GUUGCCCAUCAUCACCtg-3’) in 200 μL autoclaved distilled water was added dropwise to 0.8 mL dichloromethane containing 2.5 mg PLGA and emulsified by sonication (10% of maximum frequency for 30 s; SFX 550, Branson Ultrasonics) to form a primary W1/O emulsion. Next, 2 mL 1% (w/v) PVA1500 (Alfa Aesar) was added, and the mixture was further emulsified by sonication for 1 min to form a W1/O/W2 double emulsion. Then, 6 mL 1% (w/v) PVA1500 was added, and the dichloromethane was evaporated by magnetic stirring for 3 h at room temperature in a fume hood. Finally, the PLGA NPs were collected by centrifugation at 13,000 rpm for 10 min at 4 °C, washed twice with deionized water, and freeze-dried [21 (link), 36 (link)].
PLGA-rhodamine conjugated NPs were prepared by mixing PLGA and PLGA-rhodamine B endcapping (AV027, PolySciTech, Akina, Inc.). Nanoparticles were manufactured by mixing two types of polymers in a ratio of 9 to 1. AAV-GFP-encapsulated PLGA NPs were prepared by mixing 100 µg AAV-GFP plasmid and PLGA NPs. The NPs were diluted in double-distilled water to analyze their size and zeta potential by a dynamic light-scattering assay using the Zetasizer Nano ZS90 (Malvern Instruments), as previously described [23 (link)]. Scanning electron micrographs of the NPs were obtained with a scanning electron microscope (SNE-4500 M, SEC Co. Ltd.).
PLGA-rhodamine conjugated NPs were prepared by mixing PLGA and PLGA-rhodamine B endcapping (AV027, PolySciTech, Akina, Inc.). Nanoparticles were manufactured by mixing two types of polymers in a ratio of 9 to 1. AAV-GFP-encapsulated PLGA NPs were prepared by mixing 100 µg AAV-GFP plasmid and PLGA NPs. The NPs were diluted in double-distilled water to analyze their size and zeta potential by a dynamic light-scattering assay using the Zetasizer Nano ZS90 (Malvern Instruments), as previously described [23 (link)]. Scanning electron micrographs of the NPs were obtained with a scanning electron microscope (SNE-4500 M, SEC Co. Ltd.).
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Poly-L-lactic-acid (PLLA, Resomer, L 209 S, Evonik Industries, Essen, Germany), 1,4-dioxane (Sigma-Aldrich, Munich, Germany) and commercially available hydroxyapatite HA powder (CamCeram® III, Alhenia AG, Dättwil, Switzerland) were used for scaffold preparation.
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Acid soluble collagen type I (Coll), obtained from bovine skin (Kensey Nash Corporation DSM Biomedical, Exton, USA) and poly-L (lactic) acid (PLLA) (Lacea H.100-E, Mw = 8.4 × 104 g mol−1, PDI = 1.7, Mitsui Fine Chemicals, Dusseldorf, Germany) were used. As a solvent system, a mixture of 2,2,2-trifluoroethanol (TFE), 1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP) (Sigma-Aldrich, Staint Louis, USA) were used in a 50:50 (v/v) percentage. To crosslink Coll in the nanofibers, N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS) and ethanol (Sigma-Aldrich, Staint Louis, USA) were used. The following polymeric blend solution was used: PLLA/Coll-75/25 (w/w) prepared from a 18% (w/v) solution of PLLA and Coll dissolved in TFE:HFIP = 50:50 (v/v).
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Polyvinyl alcohol is a synthetic, water-soluble polymer. It is commonly used as a raw material in the production of various laboratory equipment and supplies.
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
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DMSO is a versatile organic solvent commonly used in laboratory settings. It has a high boiling point, low viscosity, and the ability to dissolve a wide range of polar and non-polar compounds. DMSO's core function is as a solvent, allowing for the effective dissolution and handling of various chemical substances during research and experimentation.
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Chloroform is a colorless, volatile liquid with a characteristic sweet odor. It is a commonly used solvent in a variety of laboratory applications, including extraction, purification, and sample preparation processes. Chloroform has a high density and is immiscible with water, making it a useful solvent for a range of organic compounds.
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TRIzol reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components designed for the isolation of total RNA, DNA, and proteins from a variety of biological samples. The reagent maintains the integrity of the RNA while disrupting cells and dissolving cell components.
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Dexamethasone is a synthetic glucocorticoid medication used in a variety of medical applications. It is primarily used as an anti-inflammatory and immunosuppressant agent.
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Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.
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N,N-dimethylformamide is a clear, colorless liquid organic compound with the chemical formula (CH3)2NC(O)H. It is a common laboratory solvent used in various chemical reactions and processes.
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Dichloromethane (DCM) is a colorless, volatile organic solvent. It is commonly used as a laboratory reagent and industrial solvent due to its high solvency power. DCM has a wide range of applications in various industries, including chemical synthesis, extraction processes, and cleaning.
More about "Poly-L-lactic acid"
Poly-L-lactic acid (PLLA) is a versatile, biodegradable, and biocompatible polymer derived from renewable resources like corn starch or sugarcane.
It has a wide range of applications in the biomedical and engineering fields, including tissue engineering scaffolds, drug delivery systems, and sustainable packaging materials.
PLLA exhibits excellent mechanical properties, slow degradation kinetics, and the ability to be processed into various forms like films, fibers, and 3D-printed structures.
Researchers can leverage the power of PubCompare.ai's AI-driven protocol comparison tool to quickly identify the best reproducible PLLA research protocols from literature, preprints, and patents.
This enhances efficiency and reproducibility in their studies, leading to more reliable and impactful findings.
PLLA is often used in combination with other biomaterials like polyvinyl alcohol (PVA), fetal bovine serum (FBS), and dimethyl sulfoxide (DMSO) for cell culture and tissue engineering applications.
Chloroform, TRIzol reagent, and dichloromethane (DCM) are common solvents used in PLLA processing and characterization.
Additionally, dexamethasone and penicillin/streptomycin are often incorporated to promote cell growth and prevent microbial contamination.
By utilizing PubCompare.ai's cutting-edge technology, researchers can streamline their PLLA-related studies, leading to more efficient and reproducbile research outcomes.
Explore the full capabilities of PubCompare.ai today and unlock the potential of your Poly-L-lactic acid research.
It has a wide range of applications in the biomedical and engineering fields, including tissue engineering scaffolds, drug delivery systems, and sustainable packaging materials.
PLLA exhibits excellent mechanical properties, slow degradation kinetics, and the ability to be processed into various forms like films, fibers, and 3D-printed structures.
Researchers can leverage the power of PubCompare.ai's AI-driven protocol comparison tool to quickly identify the best reproducible PLLA research protocols from literature, preprints, and patents.
This enhances efficiency and reproducibility in their studies, leading to more reliable and impactful findings.
PLLA is often used in combination with other biomaterials like polyvinyl alcohol (PVA), fetal bovine serum (FBS), and dimethyl sulfoxide (DMSO) for cell culture and tissue engineering applications.
Chloroform, TRIzol reagent, and dichloromethane (DCM) are common solvents used in PLLA processing and characterization.
Additionally, dexamethasone and penicillin/streptomycin are often incorporated to promote cell growth and prevent microbial contamination.
By utilizing PubCompare.ai's cutting-edge technology, researchers can streamline their PLLA-related studies, leading to more efficient and reproducbile research outcomes.
Explore the full capabilities of PubCompare.ai today and unlock the potential of your Poly-L-lactic acid research.